Method and apparatus for separating and concentrating a component from a multi-component material including macroparticles

• US 7,832,566 B2
• Filed: 05/25/2006
• Issued: 11/16/2010
• Est. Priority Date: 05/24/2002
• Status: Active Grant

• Current Assignee: Biomet Manufacturing, LLC (Zimmer Biomet Holdings, Inc.)
• Original Assignee: Biomet Biologics LLC (Zimmer Biomet Holdings, Inc.)
• Inventors: Swift, Matthew, Gordon, Nathan, Higgins, Joel C., Leach, Michael D.
• Primary Examiner(s): Soohoo; Tony G
• Assistant Examiner(s): MELLON, DAVID C
• Application Number:
  US11/441,275
• Publication Number:
  US 20060273049A1
• Time in Patent Office: 1,636 Days
• Field of Search: 210/782, 210/789, 210/787, 210/360.1, 210513-516, 210/518, 210/121, 210/122, 422/72, 422/101, 494/43, 494/50, 494/67, 494/16, 604/6.01, 604/6.05, 604/6.15, 604/218, 604/403
• US Class Current: 210/380.1
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: B01L 1/52 Transportable laboratories; Field kits
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0605 check valves
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B01L 3/5021 Test tubes specially adapted for centrifugation purposes centrifuges B04B5/04
• CPC Class Codes: B01L 3/50215 using a float to separate phases
• CPC Class Codes: B01L 9/54 Supports specially adapted for pipettes and burettes automated pipetting stations G01N35/10
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence

Abstract

An apparatus that allows for separating and collecting a fraction of a sample. The apparatus, when used with a centrifuge, allows for the creation of at least three fractions in the apparatus. It also provides for a new method of extracting the buffy coat phase from a whole blood sample and mesenchymal stem cells from bone reaming material. A buoy system that may include a first buoy portion and a second buoy member operably interconnected may be used to form at least three fractions from a sample during a substantially single centrifugation process. Therefore, the separation of various fractions may be substantially quick and efficient.

Claims

1. A separation system for use in a centrifuge device, comprising:

a container having a side wall sealed to and extending from a first end wall to a second end of the container, wherein the container is operable to contain a multi-component sample;

a separation system disposed in said container in a separation region of said container and near said first end wall and operable to contact said first end wall, said separation system including at least a first separation member operable to move in said separation region based at least in part on a density of said multi-component sample positionable in said separation region, said separation system further including a second separation member and a third separation member operably interconnecting said first separation member and said second separation member; and

a filtration member disposed in a filtration region of said container between said separation system and said second end of said container;

wherein said multi-component sample is operable to be positioned in said filtration region of said container from an exterior of said container and a portion of said multi-component sample is operable to be filtered by passing through said filtration member in said filtration region to said separation region of said container;

wherein said first separation member and said second separation member are connected to one another during separation of said multi-component sample;

wherein said third separation member has an internal passage that allows access to a volume between said first separation member and said second separation member;

wherein said first separation member is movable relative to said second separation member along at least a portion of a length of said third separation member.

2. The separation system of claim 1, wherein at least one of said separation members has a density less than the density of a pure mesenchymal stem cell sample and a different at least one of said separation members has a density greater than the density of said pure mesenchymal stem cell sample.

3. The separation system of claim 1, wherein said first separation member includes a density of about 1.00 g/cc to about 1.10 g/cc.

4. The separation system of claim 1, wherein said filtration member is fixed at a selected location within said filtration region of said container.

5. The separation system of claim 4, wherein said filtration member comprises at least one porous mesh filter having a mesh size of about 50 micrometers.

6. The separation system of claim 1, wherein said filtration member comprises a porous mesh having a mesh size larger than a diameter of at least one of whole blood, mesenchymal stem cells, white blood cells, platelets, or combinations thereof.

7. The separation system of claim 1, further comprising an extraction instrument operable to be interconnected with said third separation member to withdraw a selected volume through said internal passage of said third separation member from between said first separation member and said second separation member.

8. The separation system of claim 1, further comprising:

a centrifuge;

wherein said container is disposable in said centrifuge during a centrifugation process to form at least two fractions of the filtered multi-component sample in said separation region and move at least a portion of said separation system in the separation region of said container.

9. The separation system of claim 8, wherein said separation system includes said first buoy member moveable in said separation region of said container due to a force created at least in part by said centrifuge and a plunger moveable through at least a portion of said separation region with a force external to said container.

10. A separation system for use in a centrifuge device, comprising:

a container having an outer wall extending from a bottom wall and operable to contain a volume of a whole heterogenous material, including a liquid and a solid;

a filtration system positioned within said container in a filtration region for initial contact with the volume of the whole heterogeneous material, wherein said filtration system is operable to remove the solid from the liquid;

a separation system, positioned between said filtration system and said bottom wall in said container in a separation region, having a member with an outer wall that has a first face that is convex and a buoy with an outer wall that has a second face that is concave such that said first face and said second face are complementary such that said first face and said second face substantially mate to substantially eliminate any volume defined between said second face and said first face, wherein at least a portion of substantially only the liquid of the volume of the whole heterogeneous material moves sequentially through said filtration system to said separation region and is operable to be formed into fractions of the liquid; and

a fluid communication conduit extending from said separation system operable to allow extraction of only a selected fraction of the fractions from the liquid.

11. The separation system of claim 10, wherein said separation system has a specific gravity selected to depend on at least one fraction of the fractions of the liquid.

12. The separation system of claim 11, wherein said filtration system includes a porous mesh having a first average pore size supported on a support member.

13. The separation system of claim 12, wherein said support member includes openings larger than said first average pore size and supports said mesh in a single selected position in said container.

14. The separation system of claim 12, wherein said first average pore size of said mesh is about 10 micrometers to about 200 micrometers.

15. The separation system of claim 12, wherein said first average pore size of said mesh is less than about 50 micrometers.

16. The separation system of claim 10, wherein said buoy is operable to move within said container during centrifugation of said container and the volume of a whole heterogenous material, and wherein said member is a plunger operable to move relative to said buoy with an application of a force external to said container.

17. A system for separating and extracting selected fractions of a whole sample, the system comprising:

a container having a sidewall extending from a bottom wall to a top wall;

a filtration system positioned within said container near said top wall; and

a separation system positioned in said container between said filtration system and said bottom wall, said separation system having a first buoy having a first density, a second buoy having a second density, and a connection member interconnecting said first buoy and said second buoy;

wherein a selected fraction is operable to be separated between said first buoy and said second buoy;

wherein said filtration system allows for separation of a macroparticle from a fluid of the whole sample;

wherein said first buoy has an outer wall that has a first face that is convex and said second buoy has an outer wall that has a second face that is concave;

wherein said first face and said second face are complementary such that said first face and said second face substantially mate to substantially eliminate any volume defined between said first face and said second face.

18. The system of claim 17, wherein said filtration system is operable to be removed from said container and includes said top wall of said container.

19. The system of claim 17, wherein said filtration system is fixed within said container between said top wall and said separation system.

20. The system of claim 17, wherein said filtration system is in a filtration region of said container and said separation system is in a separation region of said container, wherein said filtration region is in direct fluid communication with said separation region.

21. The system of claim 17, wherein said filtration system and said separation system are positioned within said container during a centrifuge process.

22. The system of claim 17, wherein said connection member has an exterior wall and a channel wall within said exterior wall that defines a port between said first face and said second face.

23. A system for separating and extracting selected fractions of a whole sample, the system comprising:

a container having a sidewall extending from a bottom wall to a top wall;

a filtration system positioned within said container near said top wall; and

a separation system positioned in said container between said filtration system and said bottom wall, said separation system having a first buoy having a first density, a second buoy having a second density, and a connection member interconnecting said first buoy and said second buoy;

wherein a selected fraction is operable to be separated between said first buoy and said second buoy;

wherein said filtration system allows for separation of a macroparticle from a fluid of the whole sample;

wherein said second buoy is movable relative to said first buoy along at least a portion of a length of said connection member.

24. The system of claim 23, wherein said separation system, including said first buoy, said second buoy, and said connection member are all moveable within said container and relative to one another;

wherein said first density is different than said second density.