System for treating transition zone material.

(42) Date of commencement of the patent: 15. 9.1998 (45) Date of issue: 26. 4.1999

<td>(30) Priority:</td><td>(73) Patent owner:</td>

<td>10. 6.1992 US 896580 claimed. 27. 1.1993 US 009867 claimed.</td><td>PALL CORPORATION 11548 EAST HILLS (US).</td>

<td>(56) Documents:</td><td></td>

<td>WO 91 / 04088A1 WO 91 / 17809A1 US 3000540A US 4507119A US 5100564A US 5102407A US 5126054A</td><td></td>

(54) PROCESS FOR TREATING A BIOLOGICAL FLUID (57) In a method of treating a biological fluid, a buffy coat material is formed, for example, by sedimentation, then separating the transition zone material from the biological fluid, a supernatant layer, which contains platelets, and a sedimentation layer, which comprises red blood cells and finally separates the supernatant layer from the sediment layer by passing the supernatant layer through a porous medium in a downstream container.

CQ

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WR 0078018

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The present invention relates to a method for the treatment of biological fluids.

Blood consists of a number of components with different properties and uses. The separation of a single unit of donated whole blood into its components is typically facilitated by the use of differential sedimentation. The major components thus obtained are red blood cells, usually concentrated as compacted red cells (PRC), a platelet suspension, usually concentrated as platelet concentrate (PC) and plasma.

There are two main methods for separating whole blood into components. In one method, the whole blood is centrifuged to obtain a supernatant PRP fraction (platelet rich plasma) and a sedimented PRC fraction, with a transition zone material therebetween, commonly known as buffy coat, containing both leucocytes and platelets, red blood cells and plasma. The PRP fraction is separated from the buffy coat and the PRC to obtain a supernatant plasma fraction and a sedimented platelet-containing fraction. The two fractions are then separated and the platelet-containing fraction is processed to PC.

In an alternative method, the whole blood is centrifuged to obtain a supernatant platelet poor plasma fraction (PPP fraction) and a sedimented PRC fraction with a transition zone material, the buffy coat, between which contains the majority of the platelets, including leukocytes, red blood cells and Plasma. The buffy coat is separated from the supernatant PPP and the PRC sediment and centrifuged to obtain a supernatant-containing fraction and a sedimented fraction containing red blood cells. The supernatant platelet-containing fraction is then separated from the sedimented fraction and processed to form PC.

The disadvantages of the techniques include the possibility of contamination with red blood cells and / or leukocytes. With regard to the contamination with red blood cells, it should be said that the presence of red blood cells in some blood components (eg Platelet concentrate) is so undesirable that the technician operating the blood treatment devices typically constantly monitors the process during disconnection of the components and disconnects the connecting line between the blood bags when, in his opinion, as much fluid as possible has been transferred without red blood cells Passage into the downstream satellite bag was made possible. This is a labor intensive and time consuming operation.

The contamination with red blood cells creates another dichotomy. Because the platelets and plasma are valuable, blood bank personnel could try to force more of the PRP or supernatant platelet fraction into the satellite bag before stopping the flow from the collection bag. However, this is counterproductive, since the liquid squeezed out into the satellite bag may be contaminated with red blood cells, so that the squeezed liquid must be discarded or centrifuged again, which is both costly and labor intensive. Therefore, blood bank personnel could prematurely stop the flow of platelet-containing fluid before it was completely squeezed out.

In addition, the above-described techniques for the separation of whole blood into components can produce leukocyte-contaminated components. It is desirable to reduce the leukocyte concentration in each of the blood components by at least 70%, as the presence of leukocytes may adversely affect the storage time of the fractions and / or produce undesirable effects when the fractions are transfused into a patient.

In view of these problems, it may be difficult to avoid red blood cells and leukocyte contamination while maximizing the yield of the different blood components in the transition zone material or tissue. the buffy coat film. For example, the leukocyte film may be partially or completely discarded because it may be difficult to easily or efficiently separate the platelets, the plasma and the red blood cells and to deplete the leukocytes, resulting in a decreased yield of the valuable blood components such as blood cells eg Plasma and platelets, results.

The loss of platelets is particularly significant because the discarded part contains the most desirable platelets, ie the recently formed platelets. These platelets are larger and generally considered to be more active. As the younger platelets are larger, they tend to sediment more rapidly during centrifugation so that they may be concentrated at the bottom of the PRP and buffy coat by one of the techniques described above. Accordingly, because portions of the platelet-containing fluid can either be processed as part of the red blood cells or can be discarded, this represents a significant loss of the more desirable platelets.

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For example, after the PRP and PRC layers have been pressed out, the buffy coat may be discarded or processed together with the PRC layer. Similarly, after formation of a buffy coat between the PPP and PRC layers, the lower portion of the buffy coat may be processed together with the PRC layer, or the buffy coat may be partially squeezed to prevent blood platelet contamination with red blood cells.

Further, the lower portion of the supernatant containing platelets may be incompletely squeezed to prevent contamination with red blood cells from the sedimented fraction, and this may decrease the yield of platelets.

These problems are exacerbated when increasing volumes of blood components that are pooled or processed (eg, multiple units) because some of the fluid is trapped or retained in the individual collection and processing units. Overall, the small amount lost in a single assembly represents a significant loss if the high quality fluid can not be recovered.

Additionally, the processing of blood to provide blood components may result in the presence of gas or air, particularly oxygen, in the blood components or in the storage containers. This can lead to an impairment of the quality of the blood components and reduce their storage time. Further, the gas warden of air or gas in the satellite bags may pose a risk to the patient receiving a blood component by means of a transfusion.

Accordingly, the previously described methods provide a generally unsatisfactory compromise between the urgent need to maximize the yield of historically valuable blood components, such as PC, plasma, and red blood cells, from the whole blood samples while minimizing the expense and expense involved.

Therefore, there is a need for a method and system for overcoming the problems described above, which simultaneously provides maximum purity and higher yield of higher quality blood components. Further, there is a need to minimize the presence of gases. In particular, there is an urgent need for an easy-to-handle system and method of recovering and treating the transition zone material or leukocyte film which provides maximum yield and minimizes the equivalent of gases while at the same time obtaining a greater proportion of viable and physiologically active platelets.

Moreover, there is an urgent need for a method and system for efficiently combining or combining blood components, such as transition zone material or buffy coat, which maximizes the amount of fluid recoverable. Further, there is a need for a method and system that can perform combining and combining while minimizing the presence of gas.

Further, there is a need for a method and system that reduces the involvement of operators, eg, to slow down or even stop the processing of blood or blood components to prevent or minimize contamination of the desired blood components.

The present invention relates to a method for treating a biological fluid, such as blood or a blood-derived liquid product, and comprising the steps of:

Formation of a transition zone material (buffy coat) from the biological fluid, for example by sedimentation,

Separating the buffy coat from the biological fluid,

Treating the buffy coat to form a supernatant layer containing platelets and a sediment layer containing red blood cells, and

Separating the supernatant layer from the sedimented layer by passing the supernatant layer through a porous medium into a downstream container.

In describing the present invention, the following terms are used as defined below.

(A) Biological Fluid: The term biological fluid includes any treated or untreated fluid associated with a living organism, particularly blood, including whole blood, warm or cold blood, stored or fresh blood: treated blood, such as blood Blood diluted with a physiological solution, including solutions containing saline and / or anticoagulant; one or more blood components, such as Platelet concentrate (PC), platelet-rich plasma (PRP), platelet-free plasma, platelet-poor plasma (PPP), plasma, compacted red blood cells (PRC), transition zone material, buffy coat; blood analogues derived from blood or a blood component or from bone marrow: red blood cells separated from the plasma and resuspended in a physiological fluid; and platelets separated from the plasma and in

AT 405 018 B resuspended to a physiological fluid. The biological fluid may contain leukocytes or may be treated to remove leukocytes. As used herein, biological fluid refers to the components described above and similar blood products obtained in other ways and with similar properties.

As a unit is meant the amount of biological fluid obtained from a donor or derived from a unit of whole blood. The term may also refer to the amount taken during a single donation. Typically, the volume of the unit varies, with the amount varying from patient to patient and from donation to donation. Multiple units of some blood components, particularly platelets and transition zone material or buffy coat, may be pooled or combined with each other, typically combining four or more units.

(B) Transition zone material or buffy coat: The transition zone material or leukocyte film contains a red blood cell-containing material that extends across the interface between the supernatant cell-lean fraction and the sedimented cell-rich fraction of the separated biological fluid. As used herein, the boundary layer extends between the supernatant and sediment layers and includes a lower portion of the supernatant layer and an upper portion of the sediment layer as well as the material therebetween. Typically, the transition zone material or leukocyte film contains leukocytes. Preferably, the transition zone material or buffy coat may contain a high proportion of younger, more active platelets.

The transition zone material or the buffy coat may be formed by any method that separates components or fractions of blood; eg For example, the buffy coat may be formed by separation techniques based on density and / or molecular weight, eg sedimentation, more preferably centrifugation. The transition zone material or buffy coat can be formed by any centrifugation technique, including centrifugation under mild and harsh conditions. In particular, the present invention is not limited to any particular method of forming the transition zone material or the buffy coat film.

(C) Porous medium: This term describes at least one porous structure through which a biological fluid can pass. The porous medium used in conjunction with the biological fluid can be any natural or synthetic fiber, or a porous or permeable membrane (or other materials with similar surface areas and pore sizes or pore diameters) compatible with the biological fluid are (eg Blood or a blood component). The surface of the fibers or the membrane may be present in the unmodified state or modified in order to achieve a desired property. For example, the medium may be subjected to gas plasma treatment, which serves, for example, to reduce platelet adhesion.

Although the porous media may remain untreated, the fibers or membrane are preferably treated to reduce or eliminate adherence of the platelets to the media. Any treatment which reduces or eliminates platelet adhesion is considered to be within the scope of the present invention. For example, For example, the fibers may be treated on their surface as described in U.S. Patents 4,880,548, 5,100,564, and 5,152,905 to increase the critical surface wetting tension (CWST) of the fibers and reduce the adhesion tendency of the platelets. Defined in terms of CWST, a preferred range of CWST for a porous medium according to the invention is above about 70 dynes per cm, typically from about 70 dynes per cm to about 115 dynes per cm. Another preferred range is from about 90 to about 100 dynes per cm, and still another preferred range of about 93 until about. 97 dynes per cm.

A preferred range for the zeta potential (at a pH of the plasma of 7.3) is about -3 to about -30 millivolts, more preferably -7 to about -20 millivolts, and most preferably about -10 to about -14 millivolts.

The porous medium may be preformed, multilayered, and / or treated to modify the fiber surfaces, either before or after formation of the fibrous overlay. The porous medium may comprise at least one prefilter element or layer and / or a filter element layer. The porous medium may further comprise at least one element or layer to provide reinforcement, better drainage and / or improved flow properties, such as to effect a more uniform flow distribution. The porous medium may be configured in any suitable manner, such as as a flat layer, as a composite material of two or more layers, a corrugated or ribbed sheet, a woven fabric, a fibrous mat, a depth filter or a

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Membrane, although the present invention is by no means limited by this list. The porous medium may be disposed in a housing to form a fitness unit.

(D) The gap volume is the total volume of all pores within the porous medium. The void volume is hereinafter expressed as a percentage of the apparent volume of the porous medium.

&lt;E) Conversion of density when using fibers other than PBT: In the following, the term density is used and the cited density values ​​for a porous medium are based on the use of PBT fibers.

Other fibers which differ in the density of PBT may be used, provided that their surfaces by themselves or after modification have the properties mentioned above, eg a CWST greater than 70 dynes per cm.

A preferred range for the fiber diameter for practical use in this invention ranges from about 2 to 3 μm. Fiber diameters can be reported in terms of surface area, as described in U.S. Patents 4,880,548 and 5,100,564. This range is preferred because well above this range, the dimensions of the porous media and, consequently, the fluid-retaining volume of the filter assemblies become considerably larger; well below this range, the porous medium becomes relatively less coherent and is easier to squeeze.

The pore diameter of the porous medium in the context of the present invention may be determined by the modified OSUF2 method as described in U.S. Patent 4,925,572. Preferred are pore diameters which do not exceed 15 μm, more preferred are pore diameters below about 10 μm. Most preferred are pore diameter range &lt;about 6 μm.

(F) In accordance with the invention, a useful technique for determining fiber surface areas, for example with nitrogen gas adsorption, is the BET measurement method.

Fig. 1 is an embodiment of a biological care processing system comprising an assembly according to the invention; Fig. 2 is an embodiment of a processing system according to the invention; Fig. 3 is another embodiment of a biological fluid processing system according to the present invention.

Exemplary biological fluid treatment systems, which are preferably closed and / or sterile systems, are shown in the figures. As in the Rg. 1 3 and 3, the conduit system assembly 200 may include containers 20 each adapted to contain at least one unit of biological fluid, such as a fluid to receive the transition zone material or buffy coat and which are in fluid communication with a recombining device 21. In the illustrated. Embodiments, the merging device 21 includes a network or a plurality of conduits 40 leading to a single conduit 60 at an outlet or branch 50. Some of the conduits 40 have the function of inlets for the merging device 21 for the source containers 20. Alternatively, the merging device 21 may comprise a housing which includes at least two inlets and one outlet.

The outlet or branch 50 of the merging device 21 is in fuselage with a receiving or transfer container 22. In the illustrated embodiments, the flow communication with the receptacle 22 is preferably made by means of a conduit 60. Into the conduit 60, at least one device or unit is preferably interposed between the outlet or branch 50 and the receptacle 22. For example, as in the illustrated embodiments, piping assembly 200 will include a gas inlet 30, a drip chamber 31, and a gas outlet 33.

The collection or transfer container 22 may be in fluid communication with an additional container 80. In the embodiments according to FIGS. 1 and 3, the flow connection with the additional container 80 is preferably made by a line 100. Between the receiving or transfer container 22 and the additional container 80, a porous medium, such as a red cell representing a barrier medium 70 is switched.

In another embodiment of a biological fluid treatment system in accordance with the present invention, a container 90 may be in fluid communication with an additional container 80. In the embodiment according to Rg. 2 For example, the flow communication with the additional container is preferably made with a conduit 100. Between the container 90 and the additional container 80 is a porous medium, such as a medium 70, for retaining red blood cells switched.

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Embodiments of the invention may further include a gas collection section for separating gas from the flow path of the biological fluid. The gas collecting section may be designed to be suitable for a closed and / or sterile system.

For example, in the case of FIG. 3 In the illustrated embodiment, the biological fluid treatment system comprises a gas collection section 300, which preferably includes at least one conduit. The gas collection section 300 may further include a medium 150 which acts as a liquid barrier. In the illustrated embodiment, the liquid barrier forming device comprising a liquid barrier medium 150 may be interposed between the ends of the gas collection section 300, ie between lines 160 and 170.

The gas collecting section 300 may comprise a gas collecting bag (not shown), which is preferably located between the ends of the loop. From the viewpoints where the gas collecting section includes a gas collecting and displacing bag, the gas collecting and displacing bag may be used according to the invention for collecting gas and optionally for collecting a biological fluid. In one embodiment, the collected gas may be used to obtain additional biological fluid.

The gas collection section 300 may prevent mixing or contact between treated and untreated biological fluid. From these aspects of the gas collection section, which includes a liquid barrier medium 150, the gas collection section may provide assurance that biological leukocyte-contaminated fluid is isolated from leukocyte-depleted or treated biological fluid since the contaminated fluid will not pass through the fluid barrier-containing medium that sounds, can pass.

The gas collection section 300 may be in fluid communication with various components of the biological fluid processing system.

Preferably, the ends of the gas collecting section are in fluid communication with the upstream or Downstream of at least one filter unit as a barrier to red blood cells, a filter unit as a barrier to red blood cells / leukocyte depletion or a filter unit for leukocyte depletion. For example, is shown in Fig. 3 that one end of the gas collection section 300 may be connected to the conduit 100 upstream of a red blood cell barrier filter unit containing a red blood cell barrier medium 700 (connected by conduit 160) and the other end of the loop 300 may be via a Line 100 may be connected to line 100 downstream of the filter unit, which is a red blood cell barrier.

In another embodiment (not shown), the gas collection gate may be connected between the assembly 21 and the transfer container 22. For example, one end of the gas collection section may be connected to the conduit 60 downstream of the outlet 50 and the other end may be connected to the conduit 60 upstream of the transfer container 22.

Each of the components of the invention will now be described in detail below.

The containers that may be used in the biological fluid treatment unit and / or system may be made of any material and in any shape that is compatible with biological fluids and gas. A wide variety of these containers are already known in the art. For example, blood collection and satellite bags are typically made of plasticized PVC, eg PVC plasticized with dioctyl phthalate, diethylhexyl phthalate or trioctyl trimellitate. The bags may also be made of polyolefin, polyurethane, polyester or polycarbonate.

As used hereinafter, the flow connections may be made by any structure that permits the transfer of the biological fluid and / or gas from one location to another, such as, for example, the a pipe or pipe. The Flußregel- or Control devices, such as Clamps, gaskets, valves, or transfer branch shutters or the like may be disposed within or on at least one of the conduits and / or containers. The conduits used in the present invention may be made of any material that is compatible with biological fluids and gas. Preferably, they will consist of flexible material, such as Polyvinyl chloride (PVC) or plasticized PVC, eg PVC plasticized with dioctyl phthalate, diethylhexyl phthalate or trioctyl trimellitate. There may be a number of conduits which provide fluid communication with each of the individual containers, and the conduits may be arranged in the system of the present invention in a variety of ways. It is not intended to limit the present invention to a number and / or arrangement of conduits. For example, at least one conduit may be present on the side of the lid or bottom of a container or a combination thereof. At least one conduit may extend inside the container. At least one conduit may be in fluid communication with an internal passage of a container. For example, a line in

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Riessverbindung standing with the internal passage of a container which is formed by a longitudinal closure adjacent to and substantially parallel to one side of a container, wherein the termination terminates adjacent to the lower part of the container. In one aspect, the slip may flow from the lower part of the container through the internal passage to a conduit on the lid of the container.

A barrier medium for red blood cells according to the invention comprises a porous medium, which is the separation of biological cells not containing red blood cells, such as a suspension of platelets and plasma, allowed by a biological, red blood cell-containing Ruid. The red cell barrier medium prevents the biological debris containing red blood cells from entering a container such as a blood vessel a satellite bag or receptacle downstream from the barrier media. The red blood cell barrier medium can significantly reduce or effectively stop the soot of biological ruin as the red blood cell containing Ru approaches the barrier medium. For example, the red cell barrier medium may permit passage of a platelet-containing residue, but may suddenly stop the soot when red blood cells block the medium.

By reducing the soot of the biological riot, the barrier medium allows the operator to manually stop the soot to remove the biological waste containing the red blood cell from entering a container such as a container to allow a satellite bag or receptacle downstream of the barrier medium, eg before the red blood cells pass through the barrier medium. This embodiment of the invention gives the operator more time to engage and stop the soot. For example, a flow of platelet-containing fluid may be allowed to flow through the red blood cell barrier medium at an initial rate of approximately one second. 15 while the soot may decrease to about 5 ml / min as sedimented red blood cell-containing fluid approaches the medium. A reduction in soot, for example a 33% reduction, may give the operator sufficient time to stop the flow at an appropriate time. In some cases, eg When platelet-containing fluid is expressed from a plurality of separate bags at approximately the same time, this reduction in flow allows the operator to more effectively treat a larger number of containers.

A major function of the red blood cell barrier medium is the separation of a red blood cell fraction of a biological fluid from a fraction that does not contain red blood cells. The red blood cell barrier medium may act as an automatic valve by decreasing or even stopping the flow of a biological fluid containing red blood cells. In some embodiments, the automatic valve function may quickly or temporarily stop the flow of biological fluid containing red blood cells, thereby eliminating the need for the operator to observe this step.

The valve-like action is not well known, but it is believed that the flow is reduced or stopped by aggregation in or on the medium by one or more of the components of the biological riot. For example, It is presently believed that when the biological fluid containing no red blood cells passes through the medium, the leukocytes are removed from this medium. These leukocytes apparently accumulate in or on the medium, but the remainder of the non-red cell fluid typically flows through the medium.

However, if red blood cells come into direct or indirect contact with the medium, eg in direct contact with the medium or with the leukocytes, which in turn will then come into direct contact with the medium, the carbon black significantly reduced by the medium or even stopped. Without intending to be limited to a particular explanation of the mechanism of valve-like action, it is presently believed that reducing or even stopping the soot reflects aggregation of the red blood cells alone and / or in combination with leukocytes, which form a barrier which causes a prevents or blocks further flow through the porous medium. It may be that other factors, such as zeta potential, CWST and / or other properties of the fibers or porous medium may contribute to this valve-like action.

This theory for the proposed mechanism is supported by the existence of parents who are able to deplete highly efficiently leukocytes in suspensions of human red blood cells and have such small pore sizes as ca. 0.5 μm, through which red blood cells can pass freely without agglomerating, using pressures of the same order of magnitude as in the present invention. On the other hand, the filters of the present invention typically have pore diameters greater than 0.5 μm and significantly reduce or stop the flow of red blood cells when the porous medium is in contact with red cells

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Blood cells come or are penetrated by them.

In one embodiment of the invention, the efficacy of the red cell barrier medium may be increased with respect to leukocyte depletion, so that the red cell barrier medium may also function as a leukocyte depletion medium. Exemplary red blood cell barrier media and red blood cell barrier / leukocyte removal media are disclosed in US Pat. 5,100,564 and 5,152,905 and international patent application WO 91/04088.

A red blood cell barrier medium suitable for passing platelets contained in about one unit of biological fluid preferably has a fiber surface area of ​​from about 0.04 to about 3.0 meters² on, more preferably about 0.06 to 2.0 m², A preferred area for the flow area is about 3 to about 8 cm², more preferably 4 to about 6 cm², A preferred range for the relative void volume is about 71% to about 83% (corresponding to the PBT fibers at a density of about 0.23 to about 0.40 g / cm³), more preferably about 73% to about 80% (about 0.27 to about 0.37 g / cm³)

The red blood cell barrier / leukocyte depletion medium which is suitable for passing platelets from about one unit of a biological fluid preferably has a fiber surface area of ​​about 0.3 to about 2.0 meters², preferably from 0.25 to about 1.0 m², more preferably from about 0.35 to about 0.6 m² (eg 0.3 to 0.7 m²). A preferred range for the flow area is about 2.5 to about 10 cm², preferably about 3 to about 7 cm², more preferably about 3 to about 6 cm², z. B. 4 to 6 cm², A preferred range for the relative void volume is about 71% to about 83% (ie, if PBT fibers are used, corresponding to a density of the medium in the range of 0.23 to about 0.40 g / cm³), preferably about 72 to 83% (for PBT about 0.23 to about 0.35 g / cm³), more preferably about 75% to about 80%, z. B. 73 to 80% (for PBT about 0.28 to 0.35 g / cm³, z. From 0.25 to 0.33 g / cm³). The upper limits for the flow area reflect the desire to perform the filtration in a relatively short time and can be increased if longer filtration times are acceptable.

In accordance with the invention, the porous medium may be designed to remove a desired amount of leukocytes, preferably greater than 70%, more preferably greater than about 99.9 to about 99.99%, corresponding to an average residual leukocyte content per Unit less than about 0.005 x 10⁷ equivalent.

In other embodiments, which may involve different volumes of biological fluids, e.g. B. merged transition zone material, the media that serve as a barrier for red blood cells or The media serving as a red blood cell / leukocyte depletion barrier as described above may be modified as necessary. As a result, the fiber surface, the flow area, the density and the space volume can be adjusted as necessary. Z. B. For example, the fiber surface and flow area ranges listed above, which are suitable for passage of the platelets in approximately one unit of biological fluid, may be increased, e.g. B. by a factor of 6, for passing platelets of approximately 6 units of merged transition zone material or bacon skin.

Although the medium which acts as a barrier to the red blood cells may according to the invention have a substantially uniform density, another embodiment of the present invention may be constructed so that an upstream portion of the medium is of a generally lower density than a downstream portion , Z. B. For example, the density of the red blood cell barrier medium may vary continuously or stepwise while maintaining a mean average density useful for separating a supernatant layer containing platelets from a sediment layer containing red blood cells is. An exemplary medium as a barrier to red blood cells can be found in the density range in the upstream part of about 0.1 g / cm³ to about 0.23 g / cm³ lie and the density range in the discharge-side part of about 0.23 g / cm³ to about 0.40 g / cm³, In another embodiment of the invention, the medium which constitutes a red blood cell barrier may comprise two or more layers, which are preferably of different or varying density. An exemplary zonal or layered fibrous medium using PBT as the fibers may be an upstream layer having a density range of about 0.1 g / cm³ to about 0.2 g / cm³, a middle layer with a density range of 0.20 g / cm ³ to about 0.25 g / cm ³ and downstream with a density range of 0.23 g / cm³ to about 0.40 g / cm³ include.

Included within the scope of the invention are, of course, the use of other density values ​​in a particular zone or layer as well as over the entire medium containing a red

Represents blood cell barrier. These alternative density ranges can be chosen based on a desired result to be achieved, in addition to the separation of the sedimented layer from the supernatant layer, e.g. Flow rate, type of fibers used, amount

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Leukocytes that are removed, as well as other considerations.

The red blood cell barrier medium may be arranged in the system of the present invention in a variety of positions. Z. B. As shown in Figure 2, it can be arranged between two containers, such as. B. between the first container 90 and an additional container 80. As shown in Figure 1, it may downstream of the receiving or transfer container, for. B. be arranged between the receiving or transfer container 22 and an additional container 80 in the conduit 100.

A leukocyte depletion medium may, according to the invention, comprise a porous medium suitable for depleting leukocytes from the fluid flowing through the leucocyte depletion medium. A leukocyte depletion medium suitable for transmitting platelets of about one unit of biological fluid preferably has a fiber surface area of ​​from about 0.08 to about 1.0 meter² to, more preferably from about 0.1 to about 0.7 m², A preferred range for the relative gap volume is approx. 50% to approx. 89%, more preferably approx. 60% to approx. 85%. As described above in the context of the medium serving as a barrier to red blood cells, these ranges can be adapted to any need for those embodiments involving different volumes of biological fluid. Z. B .. may be increased from the fiber surface areas listed above for platelet passage of approximately 6 units of merged transition zone material or bacon skin. Exemplary media for the removal of leukocytes are described in US Pat. 5,100,564 and 4,880,548 as well as International Publication Nos. WO 91/04088.

The leukocyte-depleting medium may be arranged in the system of the invention in a variety of locations. Preferably, it is located downstream of the medium that acts as a barrier to red blood cells, ie, interposed between the medium 70, which acts as a barrier to red blood cells, and the additional container 80.

A porous medium may be used in a housing to form a filter unit. Preferably, the porous media is preformed to control dimensions, density, and pore diameters prior to assembly into the housing into a unitary, self-contained unit. Any housing of suitable shape that provides an inlet and an outlet may be used. The housing may be made of any suitable, strong, impermeable material, including an impermeable thermoplastic material, which is compatible with the fluid to be treated. The housing may include an array of one or more channels, grooves, conduits, channels, ribs, or the like, which may be serpentine, parallel, arcuate, circular, or have a variety of other configurations.

Suitable exemplary housings are disclosed in U.S. Patent Nos. 5,100,564, 5,152,905, 4,923,620, 4,880,548, and 4,925,572, as well as International Publication WO 91/04086. However, it is not intended to limit the present invention to any type, shape or structure of the housing.

The gas collection and displacement loop according to the present invention may comprise at least one conduit. It may further comprise at least one gas collection and displacement bag and a liquid barrier medium.

The loop for gas collection and displacement may contain additional elements, such. B. Flußsteuer- or -reggelgeräte as well as lines and / or connections, z. B. for connecting to the upstream and downstream side of a filter unit.

As shown in FIG. 3, the gas collection and displacement loop 300 may include first and second conduits 160 and 170 with one end of each conduit in fluid communication with the medium 150, which is a fluid barrier. Alternatively, the gas collection and displacement loop may include first and second conduits. One end of each of the conduits is in fluid communication with the gas collection and displacement bag (not shown).

A preferred gas collection and displacement loop in accordance with the present invention comprises both a liquid barrier medium and a gas collection and displacement bag with a conduit providing flow communication therebetween.

A gas collection and displacement bag is a container that is suitable for the collection and storage of gas. The gas collection and displacement bag may also be suitable for collecting and storing biological fluid. The gas collection and displacement bag may also be used to increase the recovery of biological fluid. Suitable gas collection and displacement bags include biological fluid containers previously described. In a preferred embodiment, it may be a flexible bag which can be pushed so that gas in the bag can be guided to a desired destination.

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The gas collection and displacement loop 300 may further include a liquid barrier medium 150 through which gas may pass, but not biological fluid. The medium acting as a liquid barrier may consist of various devices and elements which are suitable for separating gas present in a blood treatment system from the biological fluid being treated in the system.

A liquid barrier medium 150 comprises at least one liquid repellent, porous medium. The liquid barrier forming medium may further comprise at least one liquid compatible, porous medium. Suitable liquid-repellent, porous media and liquid-compatible media include those described in International Patent Laid-Open No. WO 91/17809 and U.S. Patent 5,126,054, but are not limited thereto. The liquid barrier medium may be enclosed in a housing to form a liquid barrier unit. Suitable housings include those disclosed in International Patent Application WO 91/17809 and US Pat. 5,126,054, but they are not limited thereto.

The conduits used in the gas collection and displacement loop may be like those described above. A flow control device as described above may be disposed within or on at least one of the conduits and / or the gas collection and displacement bag.

Typically, the gas collection and displacement loop is connected to conduits upstream and downstream of a filter unit, such as e.g. B. at least one unit that serves as a barrier to red blood cells, a unit that serves as a barrier to red blood cells / to deplete leukocytes, or a leukocyte depletion unit. The filter unit and the loop for collecting and displacing gas may be interconnected to form a single unit, such. B. a biological fluid treatment unit, and this unit may be connected to containers as desired. For example, in the embodiment illustrated in FIG. 3, the conduit 100 disposed downstream of the filter unit, the latter containing a red blood cell barrier medium 70, may be in fluid communication with a container, such as a container. B. the satellite container 80, stand. A container, such. B. the recording or the transfer container 22, which includes a platelet-containing layer, may be in fluid communication with conduit 100 upstream of the filter unit.

The arrangement of the liquid barrier medium 150 may be selected according to the desired result. Preferably, when the gas collection and displacement loop is connected at a branch to line 100 upstream of the red blood cell-forming filter unit, the latter containing a red blood cell barrier medium 70, the liquid-barrier-forming medium is brought close to the upstream branch Filter unit, which is a barrier to red blood cells are arranged. In the embodiments where a gas collection and displacement loop further comprises a gas collection and displacement bag (not shown), the bag is preferably placed adjacent to the branch downstream of the filter unit, which is a red blood cell barrier. In the embodiments comprising a gas collection and displacement loop and a red blood cell barrier / depleted leukocyte filter unit or a leukocyte depletion filter unit (not shown), at least one of the media acting as a liquid barrier and the gas collection and displacement bag is used arranged similarly with respect to the branches.

The unit for merging according to the present invention provides flow communication between at least two source containers and a receptacle, preferably between at least three source containers and a receptacle, preferably by channeling the multiple flow paths into a single flow path. As shown in FIG. 1, the merging unit 21 preferably includes a plurality of conduits 40 and an outlet or branch 50. Although there are several possibilities for the configuration of the lines, the merging device preferably comprises a network or a staggered array of lines 40, preferably with one or more branches, such as e.g. B. one or more Y compounds. As in the present case, the conduits are used to establish a flow connection between the source of biological fluid, such as a fluid. B. the separate container 20 for the units, and a container for a plurality of units to provide such. B. the transfer or receptacle 22. A flow control device may be disposed within or on at least one of the conduits.

Alternatively, the merging device 21 may comprise at least one device having a plurality of inlets and a single outlet in flow communication with a branch 50.

The device for merging as used in the present invention may be made of any material that is compatible with a biological fluid. For example, the device for merging may be made of a non-flexible material, e.g. B.

AT 405 018 Β

Acrylonitrile butadiene styrene (ABS), polycarbonate or stainless steel. Alternatively, it can be made of a flexible material, such. As polyvinyl chloride (PVC) or plasticized PVC, eg PVC plasticized with dioctyl phthalate, diethylhexyl phthalate or trioctyl trimellitate

In accordance with another embodiment of the invention, the biological fluid treatment system may include a drip chamber 31. The drip chamber 31 may be used to control the flow rate and / or to prevent gas from a container, such. B. the receiving or transfer container 22 downstream of the drip chamber, and to maximize the recovery of the biological fluid.

The drip chamber which may be used in the system may be made of any material which is compatible with the biological fluid and gas. The drip chamber can be compressible. Further, the drip chamber may contain at least one porous element, preferably a liquid repellent membrane, which allows the introduction of gas into a biological fluid treatment system and / or allows gas to be separated from a biological fluid to be treated, e.g. B. allows gas to be released from the biological fluid treatment system, but will resist or prevent the passage of a biological fluid. The porous element in this case would act as a gas inlet and / or gas outlet, as discussed later. The porous element may be arranged in a conduit or more preferably enclosed in the housing of the drip chamber. Further, the surface of the element may be oriented in various ways with respect to the flow of biological fluid. Z. B. For example, two porous elements may be disposed on opposite sides or ends of the droplet chamber or a single element within the drip chamber.

In a further embodiment of the invention, an opening, such as. B. a gas inlet and / or outlet may be associated with any of the apparatus previously described for maximizing recovery of the biological fluid in the receptacle or transfer container 22 and / or additional containers 80. Exemplary gas inlets and gas outlets and methods for their use are e.g. B. in International Publication No. WO 91/17809 and US Pat. 5,126,054.

The gas inlet 30 or the gas outlet 33 may upstream or be arranged downstream of the drip chamber 31. More preferably, as illustrated by way of example in Figure 1, the gas inlet 30 is disposed downstream of the outlet of the device 50 for merging and upstream of the drip chamber 31, which is disposed upstream of the gas outlet 33, and the gas outlet 33 is between the drip chamber 31 and the Aufnahmebzw , Transfer container 22 connected. Alternatively, a gas inlet and / or a gas outlet may be arranged in or on a drip chamber, a conduit, a filter unit or the receiving, swelling and / or additional containers.

In other embodiments (not shown), the gas inlet 30 and the gas outlet 33 may be upstream and downstream, respectively, of at least one leukocyte depletion unit, red blood cell barrier unit, or red blood cell barrier / depleted leukocyte unit be arranged.

The gas inlet and / or outlet may be arranged according to the result to be achieved, for example, according to the recovery of valuable biological fluid and / or the removal of gas, as explained in more detail below. Included within the scope of the invention is the use of more than one gas inlet and / or gas outlet.

The gas inlet is a porous element which allows gas to enter the biological fluid treatment system. Therefore, the gas inlet can provide an increase in the recovery of a valuable biological fluid (eg, transition zone material) that would otherwise be retained in various components of the system during treatment and otherwise lost.

The gas outlet is a porous element, which gas, which may be present in the biological fluid treatment system, allows it to exit the system and / or allows gas to be separated from the treated biological fluid. Thus, the gas outlet may minimize minimization of the volume of gas remaining in or in contact with the biological fluid during its treatment. The gas outlet may also allow gas to enter the biological fluid treatment system.

The gas inlet and outlet are preferably selected so that the sterility of the system is not compromised.

The gas inlet and outlet each comprise at least one porous element which is adapted to allow gas to pass therethrough. A variety of materials can be used, provided that the necessary properties of the porous element are achieved. These properties include the necessary strength in handling the pressure differences that occur in use

AT 405 018 Β and the ability to provide the desired filtration property while maintaining the desired permeability without the application of excessive pressure. The porous element of the gas inlet and the gas outlet should moreover preferably have a pore diameter of about 0.2 μm or less to preclude the penetration of bacteria into the system.

Preferably, the gas inlet and outlet include at least one liquid-repellent porous element. Since the liquid repellent porous member is not wettable or difficult to be wetted by the biological fluid to be treated in the system, the gas present in the system which encounters the liquid repellent member may pass therethrough while the biological fluid does not pass. The gas outlet may further comprise at least one liquid-compatible porous element which allows the gas to escape, but prevents it from entering the system. In a preferred embodiment of the invention, the gas outlet comprises both a liquid-repellent membrane and a liquid-compatible membrane. In addition, the gas inlet and / or the gas outlet may be enclosed in a housing which may include a lid or closure.

As already stated above, the arrangement of the gas inlet and / or the gas outlet can be selected, so that the desired result is achieved. For example, the gas inlet 30 may be located as far upstream of the conduit outlet or branch 50 as is practical to obtain sufficient maximization of recovery of the transition zone material from the assembly 200. Therefore, gas inlets may be disposed in each of the source tanks 20 of the transition zone materials to be merged. Alternatively, the gas inlet 30 may be located in a conduit 40 or downstream of the outlet or connection 50 of the merging unit 21.

In addition, it may be desirable to arrange the gas outlet 33 in the conduit 60 downstream of the outlet or linkage 50, as close as possible to the intake and delivery ports. Transfer container 22 to maximize the volume of gas removed from the conduit system assembly 200. Alternatively, the gas outlet may be disposed in the receiving or transfer container 22 and / or the container 80. The gas inlet or the gas outlet may be arranged in the drip chamber 31. In one of the embodiments of the invention, a gas inlet and / or a gas outlet between the source container 20 and the receptacle 22, for example in the line 60, be arranged.

The treatment of the biological fluid in the context of the present invention may take place at any suitable time, which may be shortly after donation. For example, when the biological fluid is dispensed as whole blood, it is typically treated as soon as practicable to maximize the number of components obtained and to maximize viability of the blood components and their physiological activity. Early treatment may reduce or eliminate more effective contamination factors, including, but not limited to, leucocytes and microaggregates. In accordance with the subject invention, the biological fluid may be treated by the donor within about 20 hours of collection. The subject invention may also include the treatment of biological fluid in accordance with practice in the United States of America wherein the treatment or processing of whole blood is generally performed within 8 hours of delivery by the donor.

The delivery of the biological fluid through the system can be achieved by maintaining a pressure differential between a container, such as a collection bag or a source container and the target of the biological fluid (eg a container such as a satellite bag or receptacle) to cause the flow to flow in a desired direction. The pressure difference can be controlled automatically or be regulated, for example as part of an automated blood treatment system or it can be manually controlled or be managed. By way of example, this pressure gradient can be achieved by a gravity gradient, the application of pressure to the collecting bag or the drip chamber (eg by hand or by means of a pressure cuff) or by placing the satellite bag in a chamber which maintains a pressure differential between the satellite bag and the collection bag (eg a vacuum chamber). Also included within the scope of the invention may be devices for expressing which produce a substantially equal pressure over the entire collection bag. It is intended that the present invention not be limited to the means for generating the pressure difference.

In general, a unit of biological fluid (eg, the donor's blood) may be transferred directly to a container, such as a collection bag, and processed to form a supernatant and a sedimented layer of the biological fluid, typically by centrifugation, so that a first supernatant layer and a first sediment layer are formed, with a

Transition zone material at the interface between the supernatant and the sedimented layer.

AT 405 018 B

The transition zone material may be separated and processed from the first overhanging layer and the sedimented layer, typically by centrifugation to form a second supernatant layer and a second sedimented layer. The second supernatant layer can be separated from the second sedimented layer. The process steps and conditions may be adapted as known to those of ordinary skill in the art. If the biological fluid is separated into a supernatant and a sedimented layer by centrifugation, the centrifugation parameters can be chosen as known to those skilled in the art. Typically, centrifugation under hard conditions at ca. 2800 xg to approx. 4800 xg for about 5 until about. 10 min performed. A typical low acceleration centrifugation may be at about 280 xg to about 1500 xg be carried out for 5 to 15 min. It is intended that the present invention is not limited to the centrifugation parameters. In a preferred embodiment, the biological fluid may be centrifuged at an increased rate of acceleration (centrifugation under harsh conditions) to form a first supernatant and a first sediment layer, as well as the transition zone material. For example, For example, whole blood can be centrifuged at high speed to form PPP and a red blood cell layer with the buffy skin extending over the interface between these layers.

In another embodiment, the biological fluid may be centrifuged at a reduced rate of acceleration (mildly-agitated) to form a first supernatant layer and a first sediment layer, as well as the transition zone material. For example, For example, whole blood may be centrifuged at a low rate to form a PRP and a red blood cell-containing layer with the buffy skin extending at the interface between these layers.

When the first protruding layer and the sedimented layer and the transition zone material are formed, the layers and / or the transition zone material may be further treated. For example, the various layers and the zone may be separated using a pressure differential to allow them to flow in a desired direction, eg from the sump to a satellite bag. Any method can be used to effect the separation.

For example, clamping the collection bag can increase the efficiency of the separation. In some embodiments, the container may be sandwiched between the first protruding layer, eg the PPP or the PRP layer, and the transition zone and / or between the first sedimented layer (eg the red blood cell layer) and the transition zone are clamped or stapled, and the single layer or the individual layers and / or the zone may flow to separate satellite bags. A similar result can be achieved with or without clamping the container, eg by a selected arrangement of the conduits with respect to the sump and its contents and / or by using a sump having an internal channel. For example, at least one conduit may be disposed on the side, top, or bottom of the container to permit separation of at least one layer and / or transition zone material from the remainder of the contents of the container through the conduit. The container may have at least one internal channel, eg a channel to the bottom of the container to provide a flow connection with a line at the top of the container and to improve the separation. Lines can be arranged on the lid and at the bottom of the container, eg for passing the supernatant and the sedimented layers from the upper and the lower part of the container. In some embodiments, at least one conduit within the interior of the container may extend to a layer or transition zone material, and the layer and / or the transition zone material may be passed through the conduit. A layer and / or a transition zone material may be passed through the conduit to a porous medium, such as a barrier medium for red blood cells.

The conduits may be selectively disconnected to allow or prevent flow into a satellite bag. For example, the operator can visually observe the bag and stop the flow by pinching off the conduit if the single layer or zone has been sufficiently squeezed out. In other embodiments, an automated blood treatment system may be used to treat the biological fluid. The automatic blood treatment system may include an observation device, eg a sensor or a photocell which observes and registers the flow rate, the pressure and / or the optical density of the expressed fluid, so that the flow can be stopped at an appropriate time. The means for generating a pressure differential may be arranged to ensure that a desired volume of fluid is squeezed out. It is intended that the invention not be limited to the manner of monitoring the flow.

The first supernatant and first sediment layer may both be squeezed, leaving only the transition zone material in the container. In another embodiment

5 405 018 Β the transition zone material is squeezed out into another container, preferably after the first supernatant layer or the first sediment layer has been pressed out. Alternatively, the transition zone material may be extruded together with a layer, eg, the first overhanging layer, and then separated.

It is intended that the present invention not be limited to the type or order of separation and / or extrusion of the layers and / or transition zone material.

Preferably, several units of biological fluid, such as Whole blood, treated in individual containers, and the resulting multiple units of transition zone material are separated and then combined, eg summarized in at least one container. The plurality of units of transition zone material may be combined or combined by any method that effects the assembly of at least two units of a biological fluid. For example, at least two containers, each containing transition zone material, are connected in series, eg vertically, so that the transition zone material is gathered in the downstream container, preferably in the lowermost container. Alternatively, two or more transition zone material containers may be simultaneously or sequentially emptied directly into a single container. In another embodiment, the plurality of units of transition zone material may be passed through a pooling unit 21 to pool or merge the transition zone material into a single container, such as, for example the receiving or transfer container 22nd

For example, in one embodiment, as shown in Figure 1, where the components are shown in a preferred vertical arrangement with the source container 20 at the highest point, the bacon skin in a plurality of containers 20 through the lines of the summarizing apparatus 21 and through the outlet or the branch 50 passed to the collection or transfer container 22nd When the bacon skin is flowing, it may at least come into contact with a device of a unit or a porous element disposed between the source container 20 and the collection or transfer container 22, eg a gas inlet 30, a drip chamber 31 or a gas outlet 33, which prevents gas from entering the receptacle and maximizing the extraction of the bacon skin.

To maximize the recovery of the transition zone material, air or gas may be introduced into the source containers 20 through the gas inlet assembly 30 or the gas outlet assembly 33, preferably using a syringe (not shown). Used in the present case, the terms air or gas refer to any gaseous fluid, such as sterilized air, oxygen, carbon dioxide and the like: it is intended that the invention is not limited to the type of gas used. While the introduced fluid is preferably ambient air or a sterile gas, some non-gaseous fluids may also be suitable. For example, For example, fluid that is lighter than and can not react with the biological fluid is included in the scope of the present invention.

The introduction of gas into the source containers 20 may be accomplished by opening a flow path from the gas inlet 30 or the gas outlet 33 to the appropriate source container 20 while closing the flow path to the collection or transfer container 22. For example, the clamps on the lines leading to the collection or transfer container 22 and all but one of the containers 20 may be closed so that when the gas is introduced into the system, the gas enters the line of the open container. In a preferred embodiment, the method includes sequentially injecting gas into the source containers 20. The flow path to each source container can be closed after the gas has been introduced into this container.

The flow path from the gas inlet 30 or the gas outlet 33 can be closed. The flow path to the first source container 20 is then opened, and when the transition zone material from the first source container 20 passes through and through the device 21 for merging in the direction of receiving or. Transfer container flows, it displaces the gas that was present in front of the column of flowing transition zone material. This gas can be expelled or removed from the system. The gas may be vented from the system through a porous element in the drip chamber or in the conduit, or preferably through an open gas outlet 33. Once the gas has been expelled from the system, the gas outlet may be inactivated to prevent gas from entering the system. For example, the gas outlet can be inactivated by manually closing the outlet, eg by closing with a cap or by clamping. Preferably, the gas outlet comprises a liquid repellent element and a liquid compatible element which automatically inactivates the outlet when wet with transition zone material.

Once the gas in front of the transition zone column has been expelled and the flow of transition zone material has stopped, the clips become adjacent to the others

Open source containers, preferably one after the other, so that transition zone material from the other

AT 405 018 Β

Containers 20 passes through the merging device 21 in the direction of the receiving or transfer container 22. The clamp, adjacent to the receiving or transfer container 22, is opened so that the transition zone material can flow into the container 22. Preferably, the clamp adjacent the receiving or transfer container 22 is opened before the clamps are opened adjacent to the other source containers.

At the beginning of the flow of transition zone material from the other source vessels, gas which is present in front of the other units of transition zone material is also displaced. Preferably, this gas is collected in the drip chamber 31 located between the outlet or branch 50 and the collection or transfer container 22. The passage of the transition zone material through the drip chamber 31, the collection of gas and / or the control or Include flow rate rules of the transition zone material.

Typically, the drip chamber 31 is inverted until the buffy skin fills the drip chamber, at which time the drip chamber is returned to its normal orientation, and the bacon skin flows to the collection or transfer container 22.

When the transition zone material flows through the system, the gas existing before the transition zone material may be expelled through the gas outlet 33 as described above. The pooled transition zone materials are then recovered in the receiving or transfer container 22, and in accordance with the invention, the introduction of air or gas into the receiving container is suppressed or minimized so that the transition zone material is recovered without collecting air.

To maximize the recovery of transition zone material, gas may be introduced behind the transition zone material retained in the system. The gas originally introduced into the source containers 20 through each of the gas inlets 30 or the gas outlets 33 follows the transition zone material as it flows through the conduits. This increases recovery of transition zone material as the gas following the transition zone material forces the fluid out of the conduits. Moreover, gas may be introduced behind the retained transition zone material by opening the gas inlet 30 after passing the transition zone material through the means for merging into the collection or transfer container 22 and collapsing the source containers 20. Additional transition zone material may then be recovered in the collection or transfer container 22.

After the recovery of the transition zone material is complete, the catchment or transfer container 22 can be sealed and separated from the system without introducing air into the container. Preferably, the collection or transfer container is heat sealed, although other methods of sealing are also suitable.

Fluid, such as Plasma, a storage solution, an additional solution or the like may be added to the transition zone material, eg during or after passing through the transition zone material into the receiving or transfer container 22. Individual units of transition zone material or transition zone material that have been combined in one container by another method can be similarly treated. If desired, the single or combined units may be stored in transition zone material for a suitable period of time. The transition zone material is separated into a supernatant and a sediment layer, typically by centrifugation, to form a second sediment layer and a second supernatant layer.

For example, in a preferred embodiment, pooled or discrete units are centrifuged on buff skin to form a supernatant and sediment layer as described above. In a more preferred embodiment, the baculocyte may be centrifuged at a low rate of acceleration (mildly-agitated centrifugation) to form a supernatant platelet-rich layer and a layer containing sedimented red blood cells.

As described in connection with the figures, can the treatment of the second supernatant layer, which typically contains a platelet-containing layer, is preferably a platelet-rich layer, the generation of a pressure difference and the conduction of the platelet-containing layer from the collecting or transfer container 22 (Figures 1 and 3) or the container 90 (Figure 2) by at least one porous medium, which in turn is at least one of the media of the red cell barrier medium, a medium as a barrier to red blood cells / depletion of leukocytes and a medium for depletion of leukocytes (not shown). Preferably, the treatment of the second supernatant layer involves separating the supernatant platelet-containing layer from the sedimented red blood cell-containing layer by passing the supernatant layer through a red blood cell barrier medium or a red blood cell barrier medium15

AT 405 018 Β chen / leukocytes is depleted until the flow through the medium slows or stops. In the embodiments illustrated in the figures, the platelet rich fluid may be recovered in an additional container 80, such as a satellite bag.

In other embodiments, a gas collection and displacement loop may be incorporated into the system and used to separate gas from a container and / or the platelet-containing fluid while maintaining a closed, sterile system. Such as described in connection with Figure 3, the platelet-containing fluid from a container, such as the receiving or transfer container 22 or the container 90, by at least one unit acting as a barrier for red blood cells, a unit that acts as a barrier to red blood cells / leukocytes depleted, or a unit that depleted leukocytes, and in a satellite container 80 are collected together with the displaced by the platelet-containing liquid gas. The gas may then be separated by means of discharges from the satellite container 80 into a gas collection and displacement loop 300.

If desired, the gas collection and displacement bag may be held in the gas collection and displacement loop above the level of the satellite bag 80 to draw the platelet-containing fluid into the satellite bag 80. When the fluid has been withdrawn, the gas collection and displacement bag may be placed lower.

The gas collection bag 80 may be squeezed to expel gas into the gas collection and displacement bag (hereinafter called gas bag) of the gas collection and displacement loop 300. For example, the satellite bag 80 may be pushed to bring the gas into the gas bag of the gas collection and displacement loop until the platelet-containing fluid reaches the gas bag. When the gas has been expelled from the satellite bag 80, the flow path between the satellite bag 80 and the gas bag in the gas collection and displacement loop 300 is preferably closed. In another embodiment of the invention, the gas and a desired amount of platelet-containing fluid may be isolated for trial purposes from the platelet-containing fluid in the satellite container 80 and collected in the gas collection and displacement loop 300. In this embodiment of the invention, the gas as described above may be ejected along with an amount of platelet-containing fluid collected in the gas bag of the gas collection and displacement loop.

After the desired amount of platelet-containing fluid has been collected in the gas bag for trial purposes, the gas can be separated from the sample by squeezing the gas bag to transfer the gas to another part of the system, eg through the barrier medium for liquids 150 of the gas collection and displacement loop, are directed into the collecting or transfer container 22. In a preferred embodiment, the gas collection and displacement loop comprises a flow control or Control device disposed between the barrier medium 150 for liquids and the connection of the loop to the line upstream of the riter unit. The flow control or Control device can manually (eg with a bracket) or automatically (eg controlled with a control valve) be managed. The platelet-containing sample may be separated or isolated at any suitable time. For example, the sample may be separated before storage or later, eg just before administration.

In another aspect, the gas collection and displacement loop may be used in conjunction with a leukocyte depletion medium to increase platelet-containing layer recovery. For example, the unit may be depleted of leukocytes between the containers, such as the collection or transfer container 22 and the satellite bag 80, and the gas can be passed into the gas bag and through the barrier medium for liquids as described above. The gas may then either enter the conduit upstream of the filter unit, after displacement into the collection or transfer container 22 or directly after passing through the liquid barrier medium, containing some platelet-containing fluid present in the filter unit and / or in the conduit downstream the riter unit is restrained, displace or can drive. This displaced, platelet-containing fluid can be recovered in the satellite bin 80 without accumulating gas since the garniture unit will not completely dry out.

Further embodiments are encompassed by the present invention. For example, with respect to the recombiner, the gas outlet may be used as a gas inlet and, conversely, the gas inlet may be used as a gas outlet at the various stages of treatment of the transition zone matrix. For example, gas may be introduced or expelled using a gas inlet and a gas outlet as described above, and the transition zone material is recovered in a collection or transfer container. The gas may then be introduced through the gas outlet so that the transition zone material remaining in the containers and / or in the filter unit 16

AT 405 018 Β unit or the filter units is retained, can be collected. Of course, gas can also be introduced through the gas inlet to a same effect.

In another embodiment, the assembly 200 may include all of the components shown in FIG. 1, except for either the gas inlet 30 or the gas outlet 33, and the assembly 200 may include a gas collection and displacement loop, eg connected to the conduit 60, downstream of the branch or outlet 50, and upstream of the collection or transfer container 22. In these embodiments, gas may enter the receiving or transfer container and the gas may be displaced from the collection or transfer container and collected in the gas collection and displacement loop.

The present invention is capable of providing a closed sterile environment. Preferably, the recovered fraction or fractions should be viable and functional, eg after a transfusion, circulate normally in the patient's bloodstream. Therefore, when the treatment has ended, the desired fraction or fractions of biological fluid can be recovered under conditions which maintain a suitable storage environment. Preferably, the fraction or fractions in a container, such as a satellite bag, and separate from the system after the container has been sealed, without the introduction of and / or impurities, particularly bacteria, into the system. Preferably, the container may be heat sealed, although other methods of sealing are also suitable.

example 1

Units of bacon skin are made and assembled by passing each unit into a bag of appropriate size. The total volume of the combined bacon skin was 345 ml.

The fitness media and housing (which form a fitness unit) will be in accordance with US Pat. 5,100,564 and International Publication WO 91/04088. The fighter unit contains a preformed red blood cell / leukocyte depletion medium and has a CWST of 93 to 97 dynes / cm, a diameter of about 6.4 cm, a flow area of ​​about 32 cm², a thickness of about 0.152 cm and a density of about 0.31 g / cm³ made of PBT fibers having a diameter of about 2.1 μm. The inlet of the unit was connected to the outlet of the satellite bag containing the aggregated bacon skins, and the outlet of the unit was connected to an empty bag suitable for contact with platelets and to form a system. The system was centrifuged to form a sedimented layer containing red blood cells and a supernatant layer of platelet-detaining fluid.

The system was carefully placed in a conventional puffing device without affecting the settled layer of red blood cells, and the assembly was prepared and the supernatant layer was squeezed out.

After 6 minutes, the red blood cell layer came into contact with the filter medium and the flow was substantially stopped. The resulting platelet-containing fluid was judged to be highly leukocyte free, that is, less than 6.9 x 10 * per unit and platelet rich, ie, 3.2 x 10 "per unit.

Example 2

A merging device used in the practice of this example may use 6 units of bacon skin in individual 60 ml single-unit containers, a 600 ml transfer container and a 600 ml satellite container, in a construction generally similar to that shown in FIG corresponds described. The assembly should be arranged substantially vertically with the merging device and the conduits leading to the transfer container with a total length of about 24 inches.

The gas inlet and outlet according to International Publication WO 91/17809 can be positioned as shown in FIG.

The filter unit may be as described in Example 1.

Clamps on the lines adjacent to the 6 single-unit containers of bacon skin as well

Clamp on the line between the transfer container and the gas outlet should be closed.

The gas outlet should be capped, and a 60 cm³ Syringe can be used to introduce air through the gas inlet and into the source containers. The piston of the syringe can be withdrawn to the 60 cm³ Brand and the cap are removed from the gas inlet, and

AT 405 018 Β then the syringe can be connected to the gas inlet. The clamp of the first unit container, which contains a unit of bacon skin, should be opened and the plunger of the syringe forward by about 5 to 10 cm³ be pressed so that air is introduced into the first single-unit container. The clamp to the first container should then be closed. The same procedure should be followed with respect to the remaining 5 unit bins.

The syringe should then be removed, the gas inlet capped again and the cap on the gas outlet removed. The clip of the first container should be opened to allow the bacon skin from the first container and the drip chamber should be turned over and pushed to fill the chamber with bacon skin. The drip chamber should be returned to its normal position and the bacon skin should flow through the drip chamber to the transfer container. Air should be expelled through the open gas outlet until the bacon skin contacts the liquid repellent membrane in the gas outlet.

At this point, the flow should be stopped and the clamp on the line leading to the transfer container should be opened and the bacon skin should flow into the transfer container.

The above-described process can be repeated for each of the remaining 5 containers. The river will be finished when the 6 containers have been emptied of bacon skin.

At this point, the remaining bacon skin should remain in the drip chamber and in the lines downstream of the gas inlet. In order to gain some of the retained bacon skin, the gas inlet should be freed from the cap and the bacon skin lines should flow into the transfer container. The tubes leading to the transfer container should be clinched and sealed hot, and the transfer container should be disconnected from the assembly unit for further treatment. The transfer container may be placed in a centrifuge tube and treated to form a sedimented fraction of red blood cells as well as a supernatant fraction containing platelets.

A filter unit may be connected to a satellite bag, and a gas collection and displacement loop may be connected upstream and downstream to the unit of riders. The gas collection and displacement loop may be connected using Y-connectors upstream and downstream of the filter unit. The gas collection and displacement loop comprises a 100 cm³ Gas collection and displacement bag and a housing containing a Bameremedium for liquids. The housing and the barrier medium for liquids form the barrier unit for liquids. The liquid barrier unit is disposed within the gas collection and displacement loop in a conduit between the Y-connector upstream of the riter unit and the gas collection and displacement bag. The liquid barrier medium comprises a liquid repellent membrane made in accordance with International Publication WO 91/17809. The barrier unit for liquids has also been prepared in accordance with International Publication WO 91/17809.

A clip may be placed on the line between the upstream side of the filter unit and the connector (hereinafter clip A) and a clip may also be placed on the line between the connector upstream of the filter unit and the liquid barrier medium (hereinafter clip B). , It may also be a clip on the line between the gas collection and displacement bag and the connector downstream of the filter unit (hereinafter referred to as clip C) and, moreover, a clip may be placed on the line between the downstream side of the filter unit and the satellite bag. This clip (hereinafter called clip D) may be located downstream of the connector connecting the downstream side of the filter unit to the gas collection and displacement loop.

The clamps A, B, C and D can be closed, and the transfer container can be connected to the line upstream of the filter unit. The filter unit can be arranged vertically. The staples A and D may be opened and the supernatant platelet-containing fluid may be squeezed out of the transfer container through the filter unit into the satellite bag until flow through the unit is automatically stopped, thereby removing the platelet-containing fraction from the sedimented fraction containing red blood cells is disconnected. The bracket A should be closed.

The gas collection and displacement bag may be raised above the level of the transfer container and the clip C should be opened. The lifting of the gas collection bag may allow additional platelet-containing fluid in the conduit downstream of the riter unit to leak into the satellite bag. After the fluid has drained, the clamp C can be closed and the gas collection and displacement bag lowered. Then the satellite bag should be compressed until the gas gathers towards the top of the satellite bag. The

AT 405 018 Β

Clamp C should then be opened while the satellite bag is still compressed to expel the gas into the gas collection and displacement bag. After the gas has been ejected, clamp C should be closed.

The tube from the outlet side of the filter unit can then be clamped and heat sealed, and the platelet-containing bag can be removed.

It is expected that more than 90% of the platelets will pass through the porous medium and that more than 99.9% of the leukocytes will be removed.

EXAMPLE 3

An assembly comprising a transfer container containing 6 units of gathered bacon skin, a filter unit, a gas collection and displacement loop (including a gas collection and displacement bag and a liquid barrier unit containing a liquid barrier medium) and a satellite bag may be assembled as in Example 2 , The brackets A, B, C and D can be arranged as in this example. The bacon skin may be centrifuged in the transfer container, and the supernatant-containing layer passes through the filter unit, and the gas in the satellite bag may be ejected as described in Example 2. However, prior to ejecting the gas from the satellite bag, the bag should be moved to mix the platelets, and when the gas has been expelled from the satellite bag, the satellite bag is compressed until the platelets enter the gas collection and displacement bag. Once the gas collection and displacement bag contains an appropriate amount of platelets, the gas collection and displacement bag may be raised until the platelets return to the satellite bag. This transfer between the satellite bag and the gas collection and displacement bag can be repeated two additional times. Then, the satellite bag can be compressed to expel the gas and a desired amount of platelets for sampling into the gas collection and displacement bag. Clamp D (downstream of the filter unit) and clamp C (on the line between the gas collection and displacement bag and the satellite bag) can be closed. The gas collection and displacement bag may be compressed until the gas collects in the upper part of the bag and then staple B located on the gas collection and displacement loop on the conduit between the connector upstream of the filter unit and the barrier medium may be opened. and the air from the gas collection and displacement bag may be pushed out of the bag by the barrier medium against liquids into the transfer container. The clamp B can then be closed and the tubes of the outlet side of the filter unit as well as the conduit leading away from the gas collection and displacement bag and from the transfer container can be clamped and heat sealed, and the satellite and gas collection and displacement bags, each of which platelets contains, can be removed. If desired, the platelet-containing fluid in the gas collection and displacement bag may serve for sampling without compromising the sterility of the platelet-containing fluid in the satellite bag.

While the invention has been described in some detail by way of illustration and example, it should be understood that the invention is susceptible to various changes and alternative forms and is not limited to the specific embodiments set forth above. It should be understood that these specific embodiments are not intended to limit the invention but, on the contrary, the invention is intended to cover all modifications, equivalents and alternatives falling within the spirit and scope of the invention.