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EP-3590556-B1 - SYSTEMS AND METHODS FOR CONCENTRATING CELLS WITH A SYRINGE AND A CENTRIFUGE

EP3590556B1EP 3590556 B1EP3590556 B1EP 3590556B1EP-3590556-B1

Inventors

  • MIN, KYUNGYOON
  • WEGENER, CHRISTOPHER J.
  • Alexander, DODGE

Dates

Publication Date
20260506
Application Date
20190625

Claims (15)

  1. A system for concentrating cells, comprising: a syringe (28) comprising a lumen (106) and an axial end (103) comprising a port (111) and a radial end (102) closed to liquid flow, wherein a plunger (108) divides the axial and radial ends, and wherein the syringe is configured to hold a cellular suspension; and a cap (104) comprising a vent (107) disposed at the radial end (102); wherein the syringe (28) is suitable for centrifugation, the plunger (108) is configured to be actuated towards the axial end (103) by air pressure being applied into the radial end (102) and the plunger (108) is configured to be actuated towards the radial end (102) by a vacuum being applied into the radial end (102), wherein the syringe has a filter (105) disposed at the radial end (102) configured to maintain sterility of the syringe (28).
  2. The system of claim 1, further comprising an actuator handle (114) configured for attachment with a radial side of the plunger (108) for actuation using a solid contact force, and wherein the filter (105) and the cap (104) are removable from the radial end (102) of the syringe (28).
  3. The system of claim 1, wherein the plunger (108) comprises a visible indicator (109) indicative of a relative position of the plunger (108) between the axial and radial ends.
  4. The system of claim 1, wherein the lumen (106) of the syringe (28) comprises a capacity of 50-60 mL.
  5. The system of claim 1, further comprising a syringe holder (116) configured to fit in a cavity of a centrifuge rotor having an axis of rotation, wherein the syringe holder (116) is configured to receive the syringe (28) containing the cellular suspension with the axial end (103) disposed towards an axial direction relative to the axis of rotation and the radial end (102) disposed towards a radial direction relative to the axis of rotation.
  6. The system of claim 1, wherein the filter (105) comprises a filter having pore sizes in the range of 0.2 to 0.45 microns.
  7. A method concentrating a cell suspension with a centrifuge, comprising: providing a syringe (28) configured to hold a cellular suspension as recited in claim 1; providing a syringe holder (116) configured to fit in a cavity of a centrifuge rotor having an axis of rotation, wherein the syringe holder (116) is configured to receive the syringe (28) containing the cellular suspension with the axial end (103) disposed towards an axial direction relative to the axis of rotation and the radial end (102) disposed towards a radial direction relative to the axis of rotation; drawing in vitro a first volume of the cell suspension into the axial end (103) of the syringe (28); centrifuging the syringe (28) containing the first volume within the syringe holder (116) with the port (111) of the axial end (103) disposed closer to the axis of rotation relative to the radial end (102) of the syringe, wherein centrifuging is performed until the first volume is separated into pelletized cells and a supernatant; expressing off the supernatant from the syringe (28) until the axial end (103) of the syringe (28) comprises the pelletized cells and a desired volume of supernatant; and resuspending the pelletized cells in the desired volume of supernatant to arrive at a final cell product.
  8. The method of claim 7, wherein the first volume of the cell suspension comprises a volume equal to or less than 50-60 mL.
  9. . The method of claim 7, further comprising providing a pneumatic pump configured for connection with the radial end (102) of the syringe (28), wherein drawing the first volume or expressing off the supernatant is performed by the pneumatic pump.
  10. The method of claim 7, further comprising: providing an actuator (114) configured for attachment with a radial side of the plunger (108); drawing the first volume by pulling the actuator (114) towards the radial end (102); and expressing off the supernatant by pushing the actuator (114) towards the axial end (103).
  11. The method of claim 7, wherein the syringe holder (116) is configured to fit into a cavity of a centrifuge rotor configured to centrifuge a 2000-mL bottle.
  12. The method of claim 7, wherein the syringe holder (116) comprises an inner shell (119) configured to fit concentrically within an outer shell (117) having an end forming a recess (121) when the inner shell (119) is fully inserted into the outer shell (117).
  13. The method of claim 7, wherein expressing off the supernatant from the syringe is performed with the port (111) of the syringe (28) disposed gravitationally above the plunger (108).
  14. The method of claim 7, wherein after the step of expressing off the supernatant and prior to resuspending the pelletized cells, a small volume of new media is drawn into the syringe (28) in preparation for resuspension to arrive at the final cell product.
  15. The method of claim 7, wherein the first volume of cell suspension comprises at least one of red blood cells, white blood cells, and platelets.

Description

Field of the Disclosure The present disclosure relates generally to systems and methods of washing biological cell suspensions and, in particular to systems and methods for washing and/or concentrating small volumes of cells with a syringe and a centrifuge. Background Many therapies and laboratory protocols currently include practices in which a targeted cellular component (e.g., red blood cells, white blood cells, platelets) is separated from a cell suspension, e.g., whole blood, and stored for later use, e.g., infusion to a patient. The targeted cellular component may be in a suspension that includes a supernatant, e.g., plasma. It may be desirable to wash the cell suspension with a wash solution, e.g., saline, to remove the supernatant as well as any nontarget cellular material prior to later use. Large volumes of wash solution are often used to clear processed fluid through fluid systems used to concentrate the target cells into final volumes ranging from approximately 50 ml to 5,000 ml. There may, however, be instances in which smaller final volumes (e.g., 10 ml or less) are desired, such as when processing single-dose quantities of blood cell products. US 2006/205581 A1 discloses a blood processing system in which a syringe is mounted in cradle which includes a shuttle that engages the piston handle of the syringe for moving the piston to draw blood into the syringe. The cradle is received in a centrifuge and, after a hard spin rotation in the centrifuge by which blood in the centrifuge is separated, the shuttle is activated to expel the separated components from the syringe into various containers that are connected to the syringe and also mounted in the cradle. Summary According to an exemplary embodiment, the present disclosure is directed to a system for concentrating cells. A syringe comprises a lumen and an axial end comprising a port and a radial end closed to liquid flow. A plunger divides the axial and radial ends, and the syringe is configured to hold a cellular suspension. A filter disposed at the radial end is configured to maintain sterility of the syringe. A cap comprises a vent disposed at the radial end. The plunger is configured to be actuated towards the axial end by air pressure being applied into the radial end and the plunger is configured to be actuated towards the radial end by a vacuum being applied into the radial end. According to an exemplary embodiment, the present disclosure is directed to a method concentrating a cell suspension with a centrifuge. Provided is a syringe comprising an axial end comprising a port and a radial end closed to liquid flow. A plunger divides the axial and radial ends, and the syringe is configured to hold a cellular suspension. Provided is a syringe holder configured to fit in a cavity of a centrifuge rotor having an axis of rotation. The syringe holder is configured to receive the syringe containing the cellular suspension with the axial end disposed towards an axial direction relative to the axis of rotation and the radial end disposed towards a radial direction relative to the axis of rotation. A first volume of the cell suspension is drawn into the axial end of the syringe. The syringe containing the first volume is centrifuged within the syringe holder with the port of the axial end disposed closer to the axis of rotation relative to the radial end of the syringe. Centrifuging is performed until the first volume is separated into pelletized cells and a supernatant. The supernatant is expressed off from the syringe until the axial end of the syringe comprises the pelletized cells and a desired volume of supernatant. The pelletized cells are resuspended in the desired volume of supernatant to arrive at a final cell product. Brief Description of the Drawings Features, aspects, and advantages of the present embodiments will become apparent from the following description, appended claims, and the accompanying exemplary embodiments shown in the drawings, which are briefly described below. Fig. 1 is a perspective view of a system for washing small volumes of cellular suspensions, according to an exemplary embodiment;Fig. 2 is a schematic view of a disposable kit for use in the system of Fig. 1, according to an exemplary embodiment;Fig. 3 is a diagrammatic illustration of a syringe, according to an exemplary embodiment;Figs. 4A and 4B are diagrammatic illustrations of a pneumatic mechanism by which the syringe of Fig. 3 is filled with fluid, according to an exemplary embodiment;Figs. 5A-5C are diagrammatic illustrations of a manual or a solid contact force mechanism by which the syringe of Fig. 3 is filled with fluid, according to an exemplary embodiment;Fig. 6 is a diagrammatic view of a fluid-filled syringe ready for centrifugation, according to an exemplary embodiment;Fig. 7A is a longitudinal cross-section view of a syringe holder suitable for use with a syringe during centrifugation, according to an exemplary embodiment;Fig. 7B is a perspective