SE469211B - SYSTEM AND PROCEDURE FOR CENTRIFUGAL SEPARATION OF TWO OR MORE FRACTIONS FROM A COMPOSITIVE SCIENCE AND THEREFORE DETERMINED PASSAGE BY Means TO STRAIGHT THE PASS TO DIVISE - Google Patents

Description

10 15 20 25 30 469 211 hrs! STATE OF THE ART In the European patent EP-B1-0 148 182 and the like U.S. Patent No. 4,617,009 discloses a system resp. a procedure and a bag which meet the above definition tioner. In the described prior art, it has been found that difficulties may arise by folding the bag within the area of the transverse throttles. Furthermore, it has proved difficult that in a conventional centrifuge with a conventional dimensional centrifuge cups and conventional types of blood clots few room with a cassette with more than one choke. One reason for this difficulty is that when using more than one choke easily get inappropriate angles for the inner cavity of the cassette or folding of the top section of the blood bag and / or the middle of the transverse throttles.

DESCRIPTION OF THE INVENTION The system of the present invention solves the above first mentioned problem in that it is provided with means for to stretch the bag laterally in the middle of said throttling or strangulation. The latter problem is solved simultaneously by a second throttling and subsequent squeezing of the bag does not need to be completely inside the used cassette. This throttling or squeezing can instead take place completely or partly after completion of centrifugation and after opening of the cassette.

Said means for stretching the bag are preferably holes provided in the bag and fixings arranged in the cassette. means for these holes, for example in the form of at the ends of the choke arranged guide studs. This achieves a very simple construction, which need not cost more either the bag or the set.

Said means for clamping suitably consists of a a pair of rails arranged parallel to the choke, e.g. described in the above-mentioned European and American patents.

UI 10 15 20 25 30 In a preferred embodiment of the system according to the finding, said pair of rails are arranged to be closed and opened with by means of a clamping device, which is arranged to enable three separate positions for the rail pair, namely a separation line fid which the pair of rails forms a narrow gap in it between the rails located the bag, a completely closed position, respectively a completely open location.

A bag suitable for the system according to the invention can consist of two mutually planned flexible foils, which along the periphery are connected to each other by a longitudinal joint, wherein at least one connecting nipple is inserted between the foils at at least one end of the elongate bag.

A simple construction for the above-mentioned clamping device is provided, if this consists of a stirrup, which has a leg rotatably mounted in one half of said pair of rails and a second leg, which is foldable over the other half of the pair of splints or part of this half, the distance between the two the legs vary in such a way that a displacement of the jumper in the longitudinal direction of the legs provides said separator ring position and the fully closed position, respectively.

The system according to the invention can in the manner described in the above-mentioned patent be provided with a second pair of rails, which is arranged to close the bag in parallel after the separation with the first pair of rails at a distance from this while the bag is still in the cassette. Alternatively, however, this can closure takes place after the bag has been removed from there. This second closure of the bag can be done either in the cassette after it has been opened or after the bag has been completely removed from the cassette ten. If the closure takes place in the latter way, it may be facilitated by a stand for hanging the bag after the separation, provided with one or more additional pairs of rails for clamping the bag parallel to the first pair of rails at a distance from this. -à 10 15 20 25 30 469 211 This stand may be provided with means for displacement of the bag in its longitudinal direction relative to said outer member. more pair of rails to enable 3 ra ï ;;; sluüning by means of these rails substantially between two separated; fractions. These funds for displacement of the bag consistently consists of a rotatable roller with fixing means for one end of the "elongated bag".

The cassette is conveniently as described above said patent in two parts and arranged to be able to be fixed together mans with the enclosed bag in a conventional pivotable centrifuge cup.

Due to the forces that occur during centrifugation, are the cassette is preferably internally provided with fixing means for locking the former pair of rails in a fixed position in relation to to the direction of separation, for example in the form of parallel to said throttling arranged grooves inside the cassette.

As mentioned above, the invention also comprises a method centrifugal separation of two or more fractions from a compound liquid, eg blood, in the case of an elongate bag filled with said liquid and placed in a cassette in a central fug and said cassette is provided with one or more relative the direction of separation transverse choking gaps, along which the bag is closed after separation. This procedure is characterized by according to the invention in that the bag is stretched in its transverse direction in the middle of the respective choke gap. The procedure according to the invention for separating different blood fractions, wherein the bag is closed along said choke gap by means of a first pair of rails after separation for separation of plasma and blood cells, then appropriate accumulated buffycoat is separated by means of of one or two additional rail pairs.

At unknown hematocrit value (cell content), appropriate two additional pairs of rails arranged parallel to the first on either side of this, the first pair being removed after that the other two pairs are closed and whereby the position of the two further the pair of rails are preferably adjusted after the separation in relation to observable separating layers between the different fractions tion. lO 15 20 25 30 * izzaa 211 ' L " The invention also includes a modified process for centrifugal sepm.eriLg of: vä or more fractions from blood, whereby the bandage ”is filled into an elongated bag, which is placed in a cassette in a reindeer carriage a transverse portion of the bag located in the middle of fdr a corresponding ¿relation to separation- the direction of transverse aperture in the cassette, this gap is closed by means of a first pair of rails after separation. the ring. This method is characterized by the bag being closed by means of one or more additional pairs of rails at the height of observable fraction limits after opening the cassette.

It has proven practical to this further sealing is performed with the bag removed from the cassette and suspended in a suitable suspension device, eg a stand provided with said additional pair of rails. The height of the pass can hereby easily adjusted in relation to said further rail pairs before closing the bag by means of this.

An opportunity to separate the different separated fractions from each other is that you start with the bag being hung first with the plasma on top, then separated by the buffycoat by means of said further pair of rails, that the bag was then turned over up and down for draining the plasma, then the plasma and the buffycoat separating the pair of rails is opened or completely removed. nas for bottling the buffycoat and that the viscous the blood cells are diluted and preferably finally transferred to a separate collection bag for these.

Preferably, however, a blood pass of the kind used is provided described in the above-mentioned patent, with hose connections in both above the subsection and combined with separate component holder, as will be described in more detail in connection with the following drawings. 10 15 20 25 469 211 When using such a bag set, the separation can available in such a way that first the buffyccat either with hi Lp 1 * two pairs of shut-off rails arranged in the cassette or with 9 LL- of two outer rail pairs are separated from the other fractions. Härefte the blood bag is hung with the plasma section at the top and one by one dilution loaded bag at a higher level than the solder bag cell concentrate. Thereafter, a fluid barrier is broken, e.g. stick, between the dilution solution and the cell concentrate. Hereby the dilution solution flows over to the cell concentrate and dilutes this. As the next step, the now empty bag is moved the dilution solution down to the lowest practical level, where the second component pass is also placed. Then a liquid barrier between this second component bag and the plasma fraction. The dilute the cell concentrate and the plasma then overflow to the respective component passes, which are suitably welded from blood pass. The shut-off rulers can then be removed and the buffycoat can spread throughout the bloodstream.

As mentioned above, the invention also includes a bag intended for use in a system of the kind defined above performing the described method, comprising two mot elongated foils, which are joined together each other along the periphery, with at least one connecting nipple being connected to the interior of the pass, preferably inserted between the two the foils at at least one end of the elongate bag.

This bag is characterized by means for stretching the bag in its transversely arranged centrally close to each other and walking edges in a suitable position in relation to the size of the fractions to be separated.

Suitably said means for stretching the bag in its transverse direction of two preferably reinforced holes arranged inside the outer edge of the pass, but outside the inner closed end of the pass space.

Ui 10 15 25 30 x i: UI l 4f; 9 221 1 Preferably, the bag according to the invention is at least wide one end provided with means for receiving in the form of holes, slots, etc. These means can first be used to fix the bag in the cassette and then also for: hanging the bag in in connection with the depletion of the various frequencies. Suitable is the bag provided with these means for hanging at both ends. This allows it to be locked in the cassette at both ends, at the same time as it can be hung after removal from the cassette partly in a first position, partly in a second position all the way up and down 'compared to the first position, for performing the above first mentioned separation procedure.

A reduced risk of confusion of the different fractions are obtained if the bag is provided with at least one connection nipple at each end.

In a preferred embodiment of the bag according to the invention said means is for stretching the bag therein transverse direction of a thickening of the pass material around for this purpose arranged holes e d.

The bag according to the invention is preferably also at least ne along its longitudinal edges provided with reinforcements in form of thickenings in the bag material. These thickeners also can be used to fix the bag in the cassette.

Suitably the bag according to the invention is included in a preferred completely closed the pass, including a supply line for the liquid to be separated, at least one outlet line for those different fractions, collection breaks for the different fractions and preferably also a bag containing diluent for the most viscous fraction, eg separated blood cells.

BRIEF DESCRIPTION OF INCLUDING DRAWINGS Fig. 1 shows an example of a blood separation phase according to the invention.

Fig. 2 shows the same bag together with a lower assembly clamping rail equipped with two locking straps.

Fig. 3 shows the bag supplemented with an upper clamping and with the stirrups swung over it. 10 15 20 25 469 211 (z w Fig. 4 shows the bag seen from the side with the squeezing the rails separated from each other. The bag is shown here with its halves filled with the liquid to be separated.

Fig. 5 shows a view corresponding to Fig. 4, but with clamping in a folded position.

Fig. 6 shows a smaller cut-out piece from the bag intended to be placed in the middle of the clamping rails.

Fig. 7 shows a section through the bag part shown in Fig. 6.

Fig. 8 shows how the bag can be divided into three parts with using an extra pair of rails.

Fig. 9 shows the bag placed in a cassette after centrifugation. ring together with a first pair of rails and one or more additional bags intended to receive the separated fractions na. The cassette is in turn placed in a conventional centrifuge galkopp.

Figures 10 and 11 show the bag centrifuged and suspended in a stand.

Fig. 12 shows a complete bag with the bag according to the find combined with additional bags intended to receive the separated fractions.

Figs. 13, 14 and 15 finally show schematically in three steps how a fractionation can be carried out using the system re- specific method according to the invention.

DETAILED DESCRIPTION OF THE SYSTEM, PROCEDURE RESPECTIVE BAG ACCORDING TO THE INVENTION The blood separation bag 1 shown in Fig. 1 is modified compared to standard bags. It is made of two on top of each other laid foils, which are joined together by means of a weld 3 a bit inside the outer edge or periphery 2. Between the periphery rin 2 and the weld 3 are at least two prepared or punched out hole 4. The bag is further provided with a slot 5 and two suspension 6. Finally, it is provided with at least two connection nipples or connecting hoses, which suitably consist of welded plastic pipes between the plastic foils. In the example shown the bag 1 is provided with four connecting nipples, namely two connection nipples 7 at one end and two connection nipples plar 7 'at the other end. 10 15 20 'i 469 9 Fig. 2 shows the bag 1 together with a squeezing rail 8, which in the following will be called the lower one The rail, In the rail are rotatably mounted two stirrups 9, which contain takes a first leg 10, which is stored in the rail 3, and a second leg 11. This second leg ll is divided into two different ones portions lla and llb at different distances from the legs 10. With 4 ' denoted two guide pins, on which the holes 4 are intended to thread ~ when the bag is placed in the cassette shown in Fig. 9.

Fig. 3 shows the bag 1 supplemented with a second assembly. clamping rail 12 and with the stirrups 9 raised above this rail 12, which will hereinafter be referred to as the upper rail.

Fig. 4 shows the bag 1 in the filled state from the side with the clamping rails 8 and 12 are shown in a separation position, ie slightly separated. The jumper 9 shown here is folded over one part of the rail 12 and inserted into a slot 13 therein.

Fig. 5 shows a view corresponding to that of Fig. 4 but with the yoke 9 offset to a pinch position. In a comparison between Figs. 2, 4 and 5 it is found that the stirrup 9 in that of Fig. 4 shown in the separation position has the part 11b in the slot 13. In the embodiment shown in fig 5, the part 11a has instead been inserted into the clamping position the slot 13 with the result that the rails 8 and 12 are clamped against each other.

Figures 6 and 7 show a plan view and a section through, respectively a smaller batch of the bag 1. Here is how to the formation of the weld 3 can cause a thickening 14, which reinforces the hole 4 and which can also extend further into the bag longitudinal direction just inside the edge 2 and form a stiffening of the bag.

Fig. 8 shows how the system according to the invention can be plated with a second pair of rails 15. This can, as shown in above-mentioned patent, be arranged inside the cassette, the bag being should have at least 4 edge holes. Alternatively, it is added only after after the bag has been removed from the cassette or at least since the cassette opened after completion of separation. 211 ' 10 15 20 25 30 469 211 10 Fig. 9 shows the bag 1 inserted in a cassette 16, as in its is inserted into a substantially conventional centrifuge cup 17.

Phase 1 is shown here after separation, but before clamping. of the sham pair 8, 12. If it is blood st: has been separated, so is part 1a filled with plasma and part 1b with blood cells. In the area 1c in between is the fraction buffjoøat. 16a denotes one inserts, which are arranged to cover at least the lower part of cassette partition plan 16b. This prevents the pass 1 wall can penetrate between the cassette halves. Further shown in Fig. 9 two weights 16c, which are arranged to press the same bag 1 at the end, which is located closest to the center of rotation. These weights 16c are suitably arranged freely movable in guides in the two the cassette halves and, during centrifugation, provide a of the bag wall.

Figures 10 and 11 show the bag 1 suspended in a stand, as in designated in its entirety 18. The phase is displayed after the separation hung on two guide pins 19 on a roller 20. Using lever e d 21, the roller can be rotated thanks to a bearing 22, which provides a raising or lowering of the bag 1. The stand post 23 supports also two rail pairs 24 and 25 respectively. For the sake of clarity, show however, Fig. 10 shows only the lower rails of the respective pair of rails and this with dashed lines. When it comes to blood separation, so contains the bag in the upper part la plasma and in the lower part lb blood cells. In the area lc on either side of the tension the rails are an accumulation of buffycoat. This can be separated by means of the rail pairs 24 and 25, which are adjustable in height.

When these pairs of rails have been clamped, the pair of rails 8, 12 can be removed. Should the buffy coat due to high or low hematocrit be completely above or below the rail pair 8, 12 as needed only one of the rail pairs 24 and 25 is used. With 26 and 27 denominations two control knobs for maneuvering the pair of rails 24 and 25. I Fig. 10 shows the pair of rails 24 and 25 separated by a spacer sleeve 28 and intended to be displaced simultaneously with this sleeve up or down. Of course, it is also possible to arrange pairs 24 and 25 for offset independently to enable the separation of both smaller and larger intermediate fractions. 1161 '. 10 15 20 25 ü 469 .Lis Fig. 12 shows a complete passport according to the invention.

Figs. 13, 14 and 15 are intended to illustrate in three steps how the pass according to Fig. 12 can be treated to separate the different blood fractions. Fig. 13 shows the leaf bag 1 provided with an anticoagulant. lance solution {to prevent blood coagulation) connected via a liquid barrier Jt eg a breaker pin 31 and hose 32 to a plasma component pass 33. At the other end of the pass 1 is via a hose 34 and a fluid barrier 35 a blood cell component bag 36 connected. Bag 36 is loaded with a dilution and nutrient solution 37 (named SAGM). As shown in Fig. 12, the bag 1 has two further connections. namely, a connection to a blood draw hose 38 with a cannula and a connection 39, which can be used to in one last moment sterile can empty buffycoat from bag 1.

In Fig. 13, blood has been drawn to the bag 1 and the hose 38 welded off. The filled blood pass 1 has, in a manner such as previously described, loaded into the cassette 16 and centrifuged to the three components plasma 40, cell concentrate 41 and buffycoat 42.

In Fig. 14, in a manner similarly described, buffycoa- trapped in space lo, the plasma in space la and the cell the center in space lb. Furthermore, bag 1 has been suspended in one stand 18 'with the plasma section laid at the top and with the bag 36 with dilution solution 37 for the cells at a higher level than the cell con- the center section lb. If the liquid barrier 35 is thereby broken, flow 37 over to the space 1b and dilute it therein. fine viscous cell concentrate.

In Fig. 15, the now empty bag 36 has been moved down to lowest practical level, where bag 33 has also been placed. also opens the liquid barrier 31 can both the plasma and that with the solution 37 dilute the cell concentrate drain into the respective component passages, which can then be welded off or otherwise separated for separate uses. Then the barrier rulers are removed, so that the buffy coat can spread throughout the bag 1 and if necessary removed through the connection 39. 211 'Jb 469 211 The invention is of course not limited to the above-described embodiments, but can be varied within the scope of the appended claims. For example, they are in the various the figures showed the details damaged very schematically. IN In practice, it may therefore be necessary in certain respects modify the same regarding saväi farm seam function.

Claims (32)

10 15 20 25 30 35 PATENT REQUIREMENTS A system for centrifugal separation of two or more fractions from a composite liquid, e.g., leaves, comprising a centrifuge having one or more cassettes (16) having an internal cavity adapted to receive an elongate bag (1) containing the liquid to be is separated, wherein at least one cassette (16) comprises one or more elongate narrowings of the inner cavity transverse to the direction of separation and means (8, 12) arranged at these throttles for squeezing the bag (1) after the separation, characterized of means (4,4 ') for stretching the bag (1) laterally in the middle of said throttling. System according to claim 1, characterized in that said means for stretching the bag (1) consists of holes (4) arranged in the bag and fixing means for these holes arranged in the cassette (16), for example in the form of guide studs arranged at the ends of the choke. (4 ') - System according to claim 1 or 2, characterized in that said means for clamping the bag (1) consists of a pair of rails (8, 12) arranged parallel to the choke. System according to claim 3, characterized in that said rail pair (8, 12) is arranged to be closed and opened by means of a clamping device (9), which is arranged to enable three separate positions for the rail pair, namely a separation position, in which the rail pair forms a narrow gap in the bag (1) located between the rails, a completely closed position and a completely open position, respectively. System according to one of the preceding claims, characterized in that the bag (1) consists of two flexible foils planned against each other, which are connected to one another along the periphery by a longitudinal joint (3), wherein at least one connecting nipple (7) is inserted between the two foils at at least one end of the elongate bag (1). * 4s9 211 * 10 15 20 25 30 35 469 211 f-l .N System according to claim 4, characterized in that said clamping device consists of a bracket (9), which has a leg (10) rotatably mounted in one half of said pair of rails (8, 12) and a second leg (11), which is foldable over the other half (12) of the rail pair (8, 12) or a part of this half, the distance between the two legs (10 and 11) varying in such a way that a displacement of the bracket (9) in the longitudinal direction of the legs provides said separation position and said completely closed position, respectively. System according to one of the preceding claims, characterized by a second pair of rails (15), which is arranged to close the bag (1) in parallel with the first pair of rails (8, 12) after the separation, either while the bag (1) is still in the cassette (16) or after it has been removed. System according to one of the preceding claims, characterized by a stand (18) for suspending the bag (1) after the separation, provided with one or more additional pairs of rails (24, 25) for clamping the bag (1) parallel to it. the first pair of rails (8,12) at a distance from this. System according to claim 8, characterized by means (19-21) for displacing the bag (1) in its longitudinal direction relative to said further rail pair (24,25) to enable a closure by means of these rails substantially between two separated fractions. System according to claim 9, characterized in that said means for displacing the bag (1) consists of a rotatable roller (20) with fixing means (19) for one end of the elongate bag (1). System according to one of the preceding claims, characterized in that the cassette (16) is two-part and arranged to be able to be fixed together with the enclosed bag (1) in a conventional pivotable centrifuge cup (17). System according to any one of claims 3-11, characterized in that the cassette (16) has internally fixing means for locking the first-mentioned pair of rails (8, 12) in a fixed position in relation to the direction of separation, for example in the form of parallel to said choke arranged grooves inside the cassette (16). 10 15 20 25 30 469 2111 '15 A system according to claim 11, comprising means (16a) for varying the volume of the inner cavity of the two-part cassette (16), characterized in that these means are arranged to at least partially follow the partition plan (run) of the cassette to prevent the bag wall penetrates between the cassette halves, System according to one of the preceding claims, characterized by means (160), which are arranged to compress the bag (1) during the centrifugation. System according to claim 14, characterized in that said means (16c) for compressing the bag (1) consists of one or more movable weights or pistons (16c) arranged closest to the center of rotation. A method for centrifugally separating two or more fractions from a composite liquid, eg blood, wherein an elongate bag (1) is filled with said liquid and placed in a cassette (16) in a centrifuge and said cassette (16) is provided with one or more throttling gaps transverse to the separation direction, along which the bag (1) is closed after the separation, characterized in that the bag (1) is stretched in its transverse direction in the middle of the throttling gap (s). A method according to claim 16 for separating different blood fractions, wherein the bag (1) is closed along said throttling gap by means of a first pair of rails (8, 12) after the separation to separate plasma and blood cells, characterized in that accumulated therebetween buffycoat is separated by one or two additional pairs of rails (l5,24,25). Method according to claim 17, characterized in that at unknown hematocrit value (cell content) two additional rail pairs (24,25) arranged in parallel with the first on either side thereof are used, the first pair (8,12) being removed after the two the second pair (24,25) is closed and the position of the two further rail pairs is preferably adjusted after the separation in relation to observable dividing layers between the different fractions. i 1 2'ï ii A method for centrifugal separation of two or fractions from blood, wherein the blood is filled on a tube placed in a cassette (16) in a centrifugal portion of the bag (1) located midway in the direction of movement transverse to this gap: the separation, one or clay drawn bag (1), led a transverse lending to separ g16) _ wherein r an equivalent in relation to cassettes first rail pair (n is closed by means of (15,24,25) at the height of observable one (16) characterized in that it (1) removed from the throttling spig of an 8,12) after with the aid characterized in bagging several additional rail pairs after the cassette fraction limits e Method according to claim 19, the closure is carried out with the bag suitable suspension device, ch suspended in a set with said further rail pair further t the cassette (16) and for example a stand (18) for characterized in that said outer (1) by means of (z4 , 2s). _ Method according to claim 20, the height position of the bag (1) is adjusted in relation to larger pairs of rails (2 this (these). A method according to any one of claims ng of a two-part cassette (16), the dividing line (16b) between the cassette halves at least by means of an insert (16a) to prevent you between the cassette halves. one of claims 13-22, characterized 4, 25) before the closure of bag 16-21, characterized in that when used, the wall of the bag (1) is partially penetrated. Method according to which the bag (1) is compressed during the non-centrifugation by means specially arranged for this purpose (16c). characterized by that A method according to claim 23, one or f said means (16c) being constituted or pistons (16c) which are arranged to press m facing the center of rotation. at the end of it, clay movable weights on the bag (1) 10 15 20 25 30 35 ”469 211 Bag intended for use in a system according to any one of claims 1-12, comprising two elongate foils facing each other, which are connected to each other along the periphery (2), wherein at least one connecting nipple (Y) is connected to the bag ( 1) inner, preferably inserted between the two foils at at least the only end of the elongate bag (1), characterized by means (4) for stretching the bag (1) in its transverse direction arranged opposite each other near the respective longitudinal edge in appropriate position in relation to the size of the fractions to be separated. Bag according to claim 25, characterized in that said means for stretching the bag in its transverse direction consists of two or more preferably reinforced holes (4) arranged inside the outer edge (2) of the bag but outside the inner closed space of the bag. Bag according to Claim 25 or 26, characterized in that it is provided at least at one end with means (5, 6, 6) for hanging in the form of holes, slots and the like. Bag according to claim 27, characterized in that it is provided with said means (5,6,6) for hanging at its two ends. Bag according to one of Claims 25 to 28, characterized in that it is provided with at least one connecting nipple (7 and 7 ', respectively) at each end. Bag according to any one of claims 25-29, characterized in that said means for stretching the bag in its transverse direction consists of a thickening (14) of the bag material around holes (4) or the like arranged for this purpose. Bag according to one of Claims 25 to 30, characterized by reinforcements (14) arranged in the form of thickenings in the bag material arranged at least along the longitudinal edges of the bag (1). Bag according to any one of claims 25-31, characterized in that it is included in a preferably completely closed bag set (1,33,36), comprising an inlet line (38) for the liquid to be separated, at least one outlet line (32, 34) for the different fractions and collection bags (33,36) for the different fractions, wherein preferably a bag (36) contains diluent (37) for the most viscous fraction, eg separated blood cells.