WO1989001792A1 - Procede et appareil de lavage de cellules - Google Patents

Procede et appareil de lavage de cellules Download PDF

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Publication number
WO1989001792A1
WO1989001792A1 PCT/US1988/002963 US8802963W WO8901792A1 WO 1989001792 A1 WO1989001792 A1 WO 1989001792A1 US 8802963 W US8802963 W US 8802963W WO 8901792 A1 WO8901792 A1 WO 8901792A1
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WO
WIPO (PCT)
Prior art keywords
bowl
fluid
supernatant
washing
cells
Prior art date
Application number
PCT/US1988/002963
Other languages
English (en)
Inventor
Richard M. Lueptow
Thomas D. Headley
Original Assignee
Haemonetics Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Haemonetics Corporation filed Critical Haemonetics Corporation
Publication of WO1989001792A1 publication Critical patent/WO1989001792A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
    • B04B—CENTRIFUGES
    • B04B5/00—Other centrifuges
    • B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
    • B04B5/0442—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3692—Washing or rinsing blood or blood constituents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
    • A61M1/3696—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M39/00—Tubes, tube connectors, tube couplings, valves, access sites or the like, specially adapted for medical use
    • A61M39/22—Valves or arrangement of valves
    • A61M39/28—Clamping means for squeezing flexible tubes, e.g. roller clamps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/02—Blood transfusion apparatus
    • A61M1/025—Means for agitating or shaking blood containers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M39/00—Tubes, tube connectors, tube couplings, valves, access sites or the like, specially adapted for medical use
    • A61M39/22—Valves or arrangement of valves
    • A61M39/223—Multiway valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
    • B04B—CENTRIFUGES
    • B04B5/00—Other centrifuges
    • B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
    • B04B5/0442—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
    • B04B2005/0464—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation with hollow or massive core in centrifuge bowl

Definitions

  • This invention is in the field of fluid processing and, more particularly, relates to the centrifugal separation and washing of contaminated body fluid, such as shed blood or glycerolized red blood cells, into two or more components, one of which is diluted less dense contaminants, i.e., supernatant, and the other of which is more dense uncontaminated desired product, i.e., red blood cells and the washing of the desired product with an appropriate solution.
  • contaminated body fluid such as shed blood or glycerolized red blood cells
  • shed blood contains whole blood, saline used to rinse the surgical site, blood clots, bone chips, fatty tissue and other miscellaneous contaminants.
  • the collected shed blood may be disposed of in at least three ways. First, it may be discarded and the lost blood volume replaced by donor blood. However, this exposes the patient to blood from donors and may result in subsequent medical risks, such as viral infection i.e., AIDS or hepatitis, or alloimmunization; an immune rejection of donor blood caused by prior sensitization by previous trans ⁇ fusions. In addition, the cost of donor blood is high.
  • viral infection i.e., AIDS or hepatitis, or alloimmunization
  • an immune rejection of donor blood caused by prior sensitization by previous trans ⁇ fusions.
  • the cost of donor blood is high.
  • a second alternative is to filter the shed blood and transfuse it to the patient.
  • the filter removes blood clots, bone chips and tissue from the shed blood, but the filtered shed blood remains diluted with the saline originally used to rinse " the surgical site. This dilution results in -about a 5% to 30% he atocrit, or red cell volume concentration, which may be an undesirable hematocrit for reinfusion to maintain patient fluid balance.
  • activated clotting factors in the supernatant of the shed blood and anticoagulants, such as heparin are not removed by filtration, and may cause subsequent medical problems when reinfused to the patient.
  • the third alternative is to "wash" the shed blood, as well as filter it.
  • centrifugal wash system One way of washing shed blood is to use a centrifugal wash system.
  • a centrifugal wash system See "The Preparation of Leukocyte-Poor Red Blood Cells: A Comparative Study, Meryman et al., Transfusion 20(3) :285:287, 1980.
  • shed blood is centrifuged while washing it with saline in a disposable centrifuge bowl or rotor.
  • a typical bowl for such a system is the so-called Latham bowl, shown in U.S. Patent No. 4,300,717. Since red cells have a higher density than saline or blood plasma, the red cells fill the outermost portion of the rotating centrifuge bowl.
  • the red cells remain in the bowl displacing the supernatant (sal ne, plasma, contaminants, etc.) out of the bowl. This concentrates the red blood cells in the bowl.
  • saline is directed into the bowl, instead of shed blood. Saline, entering the bowl, is directed by the lower extended skirt portion of the core to the outermost radius of the bowl and through the bed of packed red blood cells. In this way, the supernatant is diluted and displaced by the saline until a satisfactory "washout" efficiency is obtained.
  • washout efficiency denotes the percentage of non-red blood cell fluid, i.e., plasma and saline and contaminants, originally entering the bowl and which are removed by the wash process. A 90% or greater efficiency is typically the goal of the wash process.
  • the contents of the bowl are red cells suspended in saline.
  • the centrifugal washing procedure in conjunction with filtration concentrates the red blood cells and removes contaminants, such as blood clots, bone chips, fatty tissue and activated clotting factors. The patient can then be reinfused with his or her own washed red blood cells.
  • centrifugal wash system As above, is generally not cost-effective, unless two or more units of donor blood are needed to replace the blood shed during surgery. In two situations; surgery in which small volumes of blood are shed and post-operative blood loss, the centrifuge system has not heretofore proven cost-effective.
  • Another slower, and more labor intensive and time consuming cell washing method currently in practice is to centrifuge a unit of blood in a collection bag and subsequently manually remove the supernatant, i.e., plasma and platelets, etc., using an expressor, leaving packed red cells in the collection bag.
  • the packed red cells are diluted with saline, centrifuged again, and the supernatant manually removed, again, using an expressor, leaving packed washed red cells.
  • Another, more expensive cell washer is the IBM 2991, generally described at pages 464-465 of "Use and Analysis of Saline Washed Red Blood Cells", Wooten, Transfusion 16(5) :464:468, 1976.
  • This washer utilizes a spin and agitation method in which packed red cells are spun within a saline solution in a toroidal chamber of fixed volume and then agitated in the chamber. This process is repeated many times with fresh wash solution until sufficient hematocrit of the washed red cells is attained. The agitation is required in order to maximize interaction between the wash solution and the packed red cells.
  • This system is relatively complex and thus expensive and the procedure is time consuming because the agitation technique requires slow down and reversal of the centrifuge rotor.
  • a disposable centrifuge rotor described in parent co-pending U.S. Patent Application, Serial No. 888,764 (Attorney Docket No. HM86-03) to Thomas D. Headley entitled “Plasmapheresis Centrifuge Bowl", filed 7/22/86, is modified for use in the cell washing method of the invention.
  • the Headley rotor is generally comprised of a rotary seal and stationary header assembly, a one-piece, blow-molded integral bowl body and an optional core member.
  • the lower cylindrical separating wall portion of the core is eliminated, leaving the interior of the bowl generally radially unobstructed.
  • turbulence control means are provided to control the turbulence in the flow of fluid (supernatant or cell washing fluid) as it exits the bowl.
  • the control means allows a predetermined portion of the turbulence, created when the fluid in the rotating bowl encounters the stationary portion of the header assembly, to propagate back into the fluid in the bowl body.
  • the turbulence enhances mixing of the cell washing fluid with the supernatant and/or red blood cells.
  • the method of the invention comprises, in general, supplying unwashed body fluid, such as, unwashed shed blood or thawed glycerolized red blood cells to the lower interior of a generally radially unobstructed rigid cylindrical centrifuge bowl having separate inlet and outlet ports.
  • the fluid is supplied through an inlet port coupled through a rotary seal to a feed tube and exits at the interior bottom portion of the bowl.
  • the bowl is then rotated about its longitudinal axis.
  • the bowl rotation causes the red blood cells in the shed blood to concentrate against the inner surface of the radial periphery of the bowl.
  • the concentrated red blood cells form a first toroidal layer in an outer zone against that inner surface leaving a second inner toroidal layer of supernatant in an intermediate zone interfacing the first layer.
  • An inner coaxial zone of air consumes the rest of the space within the bowl.
  • the supply of shed blood is terminated when the observable red blood cell/supernatant interface nears a circumferential exit passageway near the rotary seal leading to an exit port in the stationary bowl header assembly. Any excess supernatant is passed out of the bowl to a waste container coupled to the exit port. While the bowl is still rotating, cell washing fluid is then admitted to the lower interior of the bowl through the same entry port in the header used to introduce the shed blood. The washing fluid displaces the supernatant toroidal layer into the waste container and replaces it with a cell washing fluid toroidal layer. Note that, unlike the prior art Latham cell washing bowl of U.S. Patent No. 4,300,717, no core skirt is available to force the wash fluid to pass through and "wash" the packed red cells in the first toroidal layer.
  • the rotational speed of the bowl is abruptly slowed, causing the two layers, i.e., the red blood cell toroidal layer and the cell washing toroidal layer, to mix together. Supernatant trapped in the concentrated red blood cells is thereby mixed with the cell washing fluid.
  • the rotational speed of the bowl is then returned to a rate at which the red blood cells are re-concentrated into the radially outward first toroidal layer, leaving a radially inner wash fluid/diluted supernatant layer.
  • a further supply of cell washing fluid is admitted to further dilute and displace the already diluted supernatant layer.
  • the bowl speed is slowed down again, to cause a further mixing and washing of the re-concentrated red blood cells with new cell washing fluid.
  • the bowl speed is again increased to pack or concentrate the red blood cells, and new cell washing fluid is introduced to displace the remaining diluted supernatant layer.
  • the rotation of the bowl is stopped, the red blood cell layer collapses and mixes with the wash solution.
  • the washed red blood cells, suspended in the wash solution, are then removed from the bowl, preferably by siphoning.
  • Fig. 1 is a partial cutaway side cross-sectional view of the centrifuge bowl of the present invention.
  • Fig. 2 is a partial cutaway sectional view of the feed tube assembly 28 of Fig. 1.
  • Fig. 3 is a bottom view of the turbulence control member taken along the lines III-III of Fig. 1.
  • Fig. 4 is a sectional view taken along the lines IV-IV of Fig. 3.
  • Fig. 5 is a top view of Fig. 4.
  • Fig. 6 is a schematic fluid/electro-mechanical diagram of the cell washing system of the present invention.
  • Fig. 7 is a partial cutaway side cross-sectional view of an alternate embodiment of the centrifuge bowl invention.
  • Fig. 8 is a bottom view of an alternate embodiment of the turbulence control member taken along the lines VIII-VIII of Fig. 7.
  • Fig. 9 is a sectional view taken along the lines IX-IX of Fig. 8.
  • the centrifuge rotor or bowl 10 comprises a seal and header assembly, shown generally at 28 of Fig. 1 and in detail in Fig. 2; a one-piece integral bowl body, shown generally at 12 (Fig. 1) ; and a turbulence control member 14 (Fig. 2) .
  • the seal and header assembly 28 is substantially identical to the seal and header assembly utilized in the Headley bowl referenced above, and incorporated herein by reference. It need not, therefore, be described in detail herein. Briefly, the assembly 28 is comprised of a non-rotatable header 30, an effluent tube 25, a feed tube assembly 24 and a rotary seal 35 formed of a seal ring 22, a flexible member 27 and an outside seal member or crown 16.
  • the header 30 has a transverse inlet bore 19 extending into a longitudinal passageway coupled to the inner bore 61 of feed tube assembly 24. Bore 61 leads to stem 18 forming an inlet fluid communication path for whole blood to enter the interior of centrifuge bowl body 12 at the lower longitudinally axial portion 12L of the bowl body. Header 30 is also provided with a transverse outlet bore 20, which extends transversely into a peripheral channel extending in parallel relationship with the feed tube assembly 24 and into an outlet coaxial passageway 62. A secondary shield 32 is formed on header 30 and extends over the rotary seal 35. A pair of complementary skirts 21 and 23 provide a passageway, therebetween, connecting the outlet passageway 62 to the top periphery of bowl body 12.
  • seal crown 16 Internal screw threads, on seal crown 16, secure the seal and header assembly to complementary threads on the bowl body.
  • a turbulence control member 14 in the shape of a circular plastic disc, is affixed to the inner peripheral wall of the ring portion 12R of bowl body 12.
  • Four circumferentially extending slots 52 are provided along the periphery of the disc 14 at the juncture between the bowl ring portion 12R and disc 14. These slots enable fluid within bowl body 12 to pass from the bowl to the pathway between skirts 21 and 23 to outlet port 20. The size and shape of these slots determine, to a large degree, the amount of turbulence propagated back to the fluid in the bowl.
  • radially indented slots 52 may be provided along the inner periphery of slots 52 to permit additional turbulence to be propagated into the bowl for mixing of wash fluid with cells»
  • a coaxial opening 50 is provided in member 14 to enable feed tube stem 18 to pass unimpeded into the lower portion 12L of the bowl body.
  • the two bowls as thus far described, are substantially the same with two exceptions.
  • the present bowl is provided with inwardly extended slots 52A and does not have a cylindrical outer wall core structure extending longitudinally and coaxial to the bowl body axis. The significance of these changes will be appreciated after a detailed description of the method of the invention.
  • fluid to be washed such as shed blood or thawed gylcerolized frozen red cells is stored in a blood reservoir 74.
  • a blood reservoir 74 As previously mentioned, for simplicity, the following description will refer to shed blood; but it should be kept in mind that other contaminated body fluids may be washed to remove contaminants, in a similar fashion.
  • Reservoir 74 and bags 26, 72 and 70 may comprise - flexible blood compatible plastic bags or rigid containers.
  • An outlet port 74A on blood reservoir 74 is connected to blood compatible tubing 76 disposed between a first clamp or valve B and connected to a first port of Y-junction 78.
  • Blood reservoir 74 is suspended above the centrifuge bowl 10 so that whole blood may be fed by gravity through tubing 76, Y-junction 78 and tubing 80 and 82 to inlet port 19 of centrifuge bowl 10.
  • wash solution bag 26 is suspended above the centrifuge bowl 10 for gravity feed coupling to inlet port 19.
  • Blood compatible tubing 83 is coupled to outlet port 26A and the tubing 83 is disposed between a second clamp S.
  • Tubing 83 is connected to a second port of Y-junction 78 coupled to tubing 80 and tubing 82 and, ultimately, to inlet port 19 of centrifuge bowl 10.
  • Outlet or effluent port 20 of centrifuge bowl 10 is coupled via blood compatible tubing 84 to inlet port 72A of waste disposal bag 72 located below bags 26 and 74.
  • Product collection bag 70 is also located below bowl 10 and connected via blood compatible tubing 86 disposed between clamp P to tubing 82 and inlet port 19 of bowl 10.
  • Product collection bag 70 is disposed below centrifuge bowl 10, so that collected product may be siphoned out the inlet port 19 of the centrifuge bowl 10, as will be described later.
  • Centrifuge bowl 10 is mounted on a chuck (not shown) for mechanical rotation about its longitudinal axis A (See Fig. 1) by motor 8 connected to bowl 10 by axle 90.
  • Motor 8 is controlled in rotational speed by motor control circuit 92.
  • bodily fluid for washing such as shed blood, or thawed glycerolized frozen whole blood
  • a wash solution such as saline, in the case of shed blood or deglycerolizing solution, in the case of glycerized whole blood, is stored m wash solution bag 26.
  • Clamps B and S are initially closed, preventing flow to bowl 10.
  • Step 1 Fill: Introduce Shed Blood
  • Motor 8 for the centrifuge bowl 10 is energized by motor control 92, and the bowl 10 is rotated until it reaches a preferred angular rotational speed, on the order of 5800 revolutions per minute.
  • the rotating centrifuge bowl is partially filled with blood from the blood reservoir 74 by opening clamp B. Clamps S and P are closed. Gravity feed from the blood reservoir 74 allows the shed blood to enter inlet port 19 and pass through the stationary header 30 and, ultimately, through the feed tube stem 18 to the lower portion of bowl body 12.
  • red blood cells are concentrated, or packed, into an outer toroidal zone 2 (shown in Fig. 1) , while supernatant is left in an intermediate toroidal zone 4 radially inwardly from the packed red blood cells 2.
  • the supernatant/red blood cell interface designated 4/2 and the supernatant/air interface designated 6/4 in Fig. 1, may be visibly seen by an operator looking down onto the top of the centrifuge bowl.
  • the supernatant is displaced out of the centrifuge bowl between skirts 21 and 23 as more blood enters the bowl and the supernatant/red blood cell interface moves radially inward.
  • the displaced supernatant exits the bowl via outlet port 20 and flows into waste disposal bag 72.
  • Step 2 Wash: Introduce Wash Solution The centrifuge bowl continues to fill with blood until the supernatant/red blood cell interface 4/2 is slightly radially outward from slots 52. After any excess supernatant has been expressed out port 20, clamp B is closed and clamp S is opened. Clamp P is left closed. This allows wash solution from bag 26 to enter the bowl through inlet port 19, to dilute and displace the remaining supernatant 4 out of the bowl via slots 52, effluent passageway 62, and outlet port 20, to inlet port 72A of waste bag 72. This leaves a radially central air zone 6, toroidal layer of supernatant 4 diluted with wash solution, and the toroidal layer of packed red blood cells 2.
  • the introduced wash solution is not forced by core skirts to enter at the lower interior bowl periphery and initially pass through the toroidal layer of packed red blood cells 2. Consequently, in the next step, a method for mixing the wash fluid and red blood cells is provided.
  • Step 3 Mix: Slow Down/Speed Up Motor
  • clamp S is closed.
  • the angular velocity of the bowl is abruptly slowed, utilizing motor control 92 to bring the rotational speed down to about 3600 RPM's.
  • This causes the two layers, 4 and 2, to intermingle and the diluted supernatant 4 to mix with the packed red blood cells 2.
  • the turbulence control slots 52A enhance the mixing of the wash solution and the supernatant by permitting turbulence to propagate into the supernatant 4. A substantial portion of any supernatant entrapped in the red cells 2 is thereby mixed with the previously diluted supernatant.
  • the speed of the bowl is increased back to its normal rotational speed of 5800 RPM, causing the red cells to reconcentrate against the inner surface of the radial periphery of the bowl 10, leaving the diluted supernatant 4 in the intermediate zone radially inwardly from the packed red blood cell layer 2.
  • Steps 2 and 3, above, may be repeated as necessary, until a satisfactory washout of supernatant is obtained by removing a sufficient amount of the initial supernatant from the red blood cells.
  • Step 4 Empty: Siphon Washed Red Cells Out of Bowl
  • the motor 8 is braked by motor control 92 and the washed red blood cells become suspended in the wash solution. Clamps B and S remain closed, while P is opened, to permit the suspended washed red blood cells to be siphoned from the bowl 10.
  • the siphon is primed by pressing the side walls of waste bag 72, which causes the fluid in bowl 10 to siphon out the port 19 to tubes 82 and 86 through open clamp P to inlet port 70A into product bag 70.
  • Tests were performed, in accordance with the above method, using donor blood diluted with dyed saline to the hematocrit typical of shed blood, i.e., 20% hematocrit.
  • a bowl speed of 5800 RPM was employed for cell concentration.
  • Step 1 the initial concentration phase (Step 1) , about 60% of the dye was removed.
  • Steps 2 to 3 four times between 5800 RPM and 3600 RPM (saline flow rate: lOOml/min; 2 minute saline flow on first cycle; 1 minute saline flow on subsequent cycles) removed an additional 30% of the original dye.
  • saline flow rate lOOml/min; 2 minute saline flow on first cycle; 1 minute saline flow on subsequent cycles
  • the system above described overcomes many disadvantages of the prior art.
  • the system incorporates a modified low-cost blow-molded bowl and an insertable molded control member.
  • This bowl and member is inherently less expensive to manufacture, since the bowl body 12 and turbulence control member 14 consists of two easily molded pieces, instead of the four complex molded pieces.
  • the system does not require the precise process control necessary for the prior art Latham bowl. This is because the fluid path in the bowl does not narrow at the top of the bowl, so red cell spillover is unlikely. Consequently, the apparatus used in conjunction with the bowl does not require expensive optical sensors and microprocessor controls, or a trained operator.
  • a simple gravity-fed flow system can be used instead of pumps and valves, because exact control of the flow to prevent spillover is unnecessary. This further reduces the machine and disposable cost and increases ease of use.
  • the fluid flow during the wash procedure is also distinctly different in the Latham bowl, as compared to the flow in the modified blow-molded Headley bowl, described herein.
  • the wash solution flows to the outermost radius and through the bed of packed red cells. As the wash solution percolates through the packed red cells, contaminants trapped in the bed of packed red cells are removed.
  • the wash solution is not initially directed through the bed of packed red cells. Instead, it flows through the supernatant along the radially inward zone of the bowl, as indicated by the arrows in Fig. 1.
  • the present invention enables use of the low cost Headley bowl by providing the above described slow-down/speed-up protocol to wash out the contaminants trapped in the packed red cell bed zone.
  • the fluid in the bowl tends to maintain its rotational momentum as the bowl slows.
  • the action of viscous forces between the fluid and the bowl, each rotating at a different speed slows the fluid to the bowl's speed. If the speed change is abrupt, the viscous forces generate turbulence and mixing in the bowl. It should be noted that this procedure differs from the prior art agitation system, referenced above, which relies on reversal of rotation to produce agitation, as in a conventional clothes washer.
  • the present invention depends on mixing produced by differential fluid rotation as the bowl slows down and speeds up more quickly than the fluid. This can be accomplished quickly, since the bowl does not need to come to a complete stop and reverse directions, as it does in an agitation cycle.
  • An alternate skirt embodiment of the invention will now be described in connection with Figs. 7-9, wherein like parts shown in previous figures carry the same reference numerals and unlike parts carry a prime suffix.
  • the centrifuge slows and then speeds up to mix the red blood cells and supernatant.
  • the violent motion of the fluid during the mix cycle permits red blood cells to "splash" through the fluid exit slots 52 into the header region.
  • these red blood cells are washed out of the bowl with the waste supernatant.
  • red blood cells results in two problems. First, the spillage is repeated with each mix/wash cycle. In the course of a typical protocol consisting of four mix/wash cycles, it is estimated that as much as 1.0 percent of the red blood cells can be lost by spillage. Since the goal of cell saving is to salvage red blood cells shed in surgery, it is desirable to lose as few red blood cells as possible. This problem is accentuated when the operator inadvertently fills the bowl with too many red blood cells. This drives the red blood cell/supernatant interface radially inward and results in even more red blood cells spilling into the header.
  • the second problem is involved with the observed color of the fluid exiting the bowl during the wash cycle following a mix cycle.
  • the supernatant of the shed blood contains free hemoglobin that has been liberated from red cells damaged during surgery.
  • the hemoglobin colors the supernatant red.
  • the red color of the supernatant diminishes as the blood is washed.
  • the operator will use the visual change of the color of the supernatant from red to clear to subjectively determine the effectiveness of the wash cycle.
  • red cells are present in the supernatant, the color change can be masked making visual detection of washout difficult.
  • these problems are avoided by using a turbulence control member 14' modified to include a short longitudinally extending cylindrical wall skirt 15. The remainder of the bowl 10' is identical to that previously described and that description will therefore not be repeated.
  • the member 14' is also identical to member 14, previously described, except for the cylindrical wall, or skirt, 15 that extends axially into the bowl from the periphery of the turbulence control member 14* . Note that the skirt 15 is disposed radially outward from the fluid exit slots 52.
  • the function of the skirt is shown in Fig. 7.
  • the bowl 10• is filled with shed blood until the red blood cell/supernatant interface 4/2 is just radially outside of the skirt 15. At this point, only supernatant is in contact with the fluid exit slots 52.
  • the red blood cells 2 and the supernatant 4 mix throughout most of the bowl.
  • the supernatant radially inside the skirt does not readily mix with red cells because of the shielding provided by the skirt. Thus, only supernatant remains in contact with the fluid exit slots 52. If the skirt were not present, supernatant and red blood cells would mix in the region just below the fluid exit slots. As a result, red blood cells could pass through the fluid exit slots and into the header region.
  • the effect of the skirt 15 is to prevent red blood cell spillage caused by splashing red cells in the fluid exit slots during the mix cycle and subsequent washing.
  • the skirt simply allowing the red blood cells caught in the header region to sediment back into the bed of packed red blood cells would not solve this problem. This is because the red blood cells that are splashed into the header region during the mix are forced into the effluent tube by pressure changes in the bowl. Once red blood cells are in the non-rotating effluent tube, they will not sediment. Only by preventing, red blood cells from reaching the fluid exit slots will red blood cell spillage be prevented.
  • the skirt accomplishes this desirable result in a simple and effective manner.
  • the bowl 10, product bag 70, and tubing 86, 84, 80, 83 and 76 will be fabricated as an entity with coupling members, such as bag spikes, affixed to the ends of the unattached tubing 83, 76 and 84.
  • the assembled unit as above described, can be sterilized and supplied as a disposable unit for subsequent attachment by the end user to a wash solution container, blood reservoir or waste container.
  • solenoid valves or hydraulic valves may be employed, other type centrifuge bowls may be used, and pumps, instead of gravity feed, employed.
  • Sophisticated monitoring and control techniques may also be employed to activate and monitor the process.

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  • Health & Medical Sciences (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Vascular Medicine (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Anesthesiology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
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  • Centrifugal Separators (AREA)

Abstract

Procédé et appareil permettant de concentrer des globules rouges et de les laver, afin de retirer le surnageant, dans lesquels on centrifuge du sang dans une cuvette de centrifugation intégrale sans noyau, à une vitesse prédéterminée, afin de comprimer les globules rouges les plus denses, laissant une couche toroïdale de surnageant moins dense. Le fluide de lavage de cellules évince le surnageant de la cuvette. Un cycle de mélange est établi, dans lequel la centrifugation est ralentie pour mélanger le fluide de lavage et les globules rouges. Ensuite on accélère à nouveau la centrifugation puis on ajoute du fluide de lavage. Le cycle de mélange est successivement répété jusqu'à obtention d'un nettoyage satisfaisant.
PCT/US1988/002963 1987-08-28 1988-08-23 Procede et appareil de lavage de cellules WO1989001792A1 (fr)

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US9032587A 1987-08-28 1987-08-28
US090,325 1987-08-28

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WO1989001792A1 true WO1989001792A1 (fr) 1989-03-09

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2668714A1 (fr) * 1990-11-05 1992-05-07 Cobe Lab Lavage des cellules.
WO1994021311A2 (fr) * 1993-03-24 1994-09-29 Haemonetics Corporation Systeme de mesure du volume dans un reservoir de sang
US5445593A (en) * 1992-08-14 1995-08-29 Fresenius Ag Method and apparatus for the continuous conditioning of a cell suspension
US5478479A (en) * 1994-05-20 1995-12-26 Haemonetics Corporation Two-stage cell wash process controlled by optical sensor
US5643193A (en) * 1995-12-13 1997-07-01 Haemonetics Corporation Apparatus for collection washing and reinfusion of shed blood
DE19802321C2 (de) * 1998-01-23 2000-05-11 Fresenius Ag Verfahren und Vorrichtung zur Aufbereitung von intra- oder postoperativen Blutverlusten für die Autotransfusion
WO2017011527A1 (fr) 2015-07-13 2017-01-19 Haemonetics Corporation Système et procédé permettant de retirer la graisse de sang récupéré
US10500330B2 (en) 2014-10-07 2019-12-10 Haemonetics Corporation System and method for washing shed blood
US11541161B2 (en) 2016-06-24 2023-01-03 Haemonetics Corporation System and method for continuous flow red blood cell washing

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US3982691A (en) * 1974-10-09 1976-09-28 Schlutz Charles A Centrifuge separation and washing device and method
FR2368966A1 (fr) * 1976-11-01 1978-05-26 Union Carbide Corp Procede de lavage du sang
FR2368965A1 (fr) * 1976-11-01 1978-05-26 Union Carbide Corp Appareil de lavage du sang
US4300717A (en) * 1979-04-02 1981-11-17 Haemonetics Corporation Rotary centrifuge seal
SE437331B (sv) * 1976-10-06 1985-02-25 Haemonetics Corp Plasmaferesapparat

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US3982691A (en) * 1974-10-09 1976-09-28 Schlutz Charles A Centrifuge separation and washing device and method
SE437331B (sv) * 1976-10-06 1985-02-25 Haemonetics Corp Plasmaferesapparat
FR2368966A1 (fr) * 1976-11-01 1978-05-26 Union Carbide Corp Procede de lavage du sang
FR2368965A1 (fr) * 1976-11-01 1978-05-26 Union Carbide Corp Appareil de lavage du sang
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TRANSFUSION, September-October 1976, Volume 16, Number 5, M.J. O'CONNOR WOOTEN, "Use and Analysis of Saline Washed Red Blood Cells", Methods. *

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2668714A1 (fr) * 1990-11-05 1992-05-07 Cobe Lab Lavage des cellules.
US5445593A (en) * 1992-08-14 1995-08-29 Fresenius Ag Method and apparatus for the continuous conditioning of a cell suspension
US5607830A (en) * 1992-08-14 1997-03-04 Fresenius Ag Method for the continuous conditioning of a cell suspension
WO1994021311A2 (fr) * 1993-03-24 1994-09-29 Haemonetics Corporation Systeme de mesure du volume dans un reservoir de sang
WO1994021311A3 (fr) * 1993-03-24 1994-11-10 Haemonetics Corp Systeme de mesure du volume dans un reservoir de sang
US5458566A (en) * 1993-03-24 1995-10-17 Haemonetics, Inc. Reservoir volume sensing systems for autologous blood recovery
EP1166806A3 (fr) * 1993-03-24 2003-10-08 Haemonetics Corporation Système de mesure du volume d'un réservoir de sang
US5478479A (en) * 1994-05-20 1995-12-26 Haemonetics Corporation Two-stage cell wash process controlled by optical sensor
US5643193A (en) * 1995-12-13 1997-07-01 Haemonetics Corporation Apparatus for collection washing and reinfusion of shed blood
US5971948A (en) * 1995-12-13 1999-10-26 Haemonetics Corporation Apparatus for collection, washing, and reinfusion of shed blood
US6299784B1 (en) 1998-01-23 2001-10-09 Fresenius Ag Method and apparatus for processing intra- or postoperative blood loss for autotransfusion
DE19802321C2 (de) * 1998-01-23 2000-05-11 Fresenius Ag Verfahren und Vorrichtung zur Aufbereitung von intra- oder postoperativen Blutverlusten für die Autotransfusion
US6814862B2 (en) 1998-01-23 2004-11-09 Fresenius Ag Method and apparatus for processing intra or postoperative blood loss for autotransfusion
US10500330B2 (en) 2014-10-07 2019-12-10 Haemonetics Corporation System and method for washing shed blood
WO2017011527A1 (fr) 2015-07-13 2017-01-19 Haemonetics Corporation Système et procédé permettant de retirer la graisse de sang récupéré
CN107735120A (zh) * 2015-07-13 2018-02-23 美国血液技术公司 用于从回收的血液中清除脂肪的系统和方法
JP2018526046A (ja) * 2015-07-13 2018-09-13 ヘモネティクス・コーポレーションＨａｅｍｏｎｅｔｉｃｓ Ｃｏｒｐｏｒａｔｉｏｎ 回収血液から脂肪を除去するためのシステム及び方法
EP3322459A4 (fr) * 2015-07-13 2019-03-13 Haemonetics Corporation Système et procédé permettant de retirer la graisse de sang récupéré
US10668207B2 (en) 2015-07-13 2020-06-02 Haemonetics Corporation System and method for removing fat from salvaged blood
JP2021120029A (ja) * 2015-07-13 2021-08-19 ヘモネティクス・コーポレーションHaemonetics Corporation 回収血液から脂肪を除去するためのシステム及び方法
US11541161B2 (en) 2016-06-24 2023-01-03 Haemonetics Corporation System and method for continuous flow red blood cell washing

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