System and process for separating a material

• US 8,337,711 B2
• Filed: 02/27/2009
• Issued: 12/25/2012
• Est. Priority Date: 02/29/2008
• Status: Active Grant

• Current Assignee: Biomet Manufacturing, LLC (Zimmer Biomet Holdings, Inc.)
• Original Assignee: Biomet Biologics LLC (Zimmer Biomet Holdings, Inc.)
• Inventors: Dorian, Randel, Leach, Michael D., Storrs, Richard W., Chavarria, Jason
• Primary Examiner(s): Reifsnyder, David A
• Application Number:
  US12/395,085
• Publication Number:
  US 20090221075A1
• Time in Patent Office: 1,397 Days
• Field of Search: 210/782, 210/789, 210/360.1, 210/380.1, 210/516, 422/72, 422/527, 604/6.01, 604/6.15, 604/403
• US Class Current: 210/782
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 3/5021 Test tubes specially adapted for centrifugation purposes centrifuges B04B5/04
• CPC Class Codes: B01L 3/50825 Closing or opening means, corks, bungs closures for containers B65D; means for removing stoppers B67B7/02
• CPC Class Codes: B01L 3/5635 connecting two containers face to face, e.g. comprising a filter
• CPC Class Codes: C12M 47/02 Separating microorganisms from the culture medium; Concentration of biomass separating microorganisms from their culture media C12N1/02
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 1/286 involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising microtomes G01N1/06; pulverising in general B02C; mixing in general B01F
• CPC Class Codes: G01N 1/4077 by other techniques involving separation of suspended solids

Abstract

Disclosed is a system to separate, enrich, and/or purify a cellular population from a biological tissue, such as a tissue sample. For example, an adipose tissue sample can be acquired and disrupted. The disrupted tissue sample can then be separated and purified. The separated components can include multipotent, pluripotent, or other cell populations.

Claims

1. A method of separating a selected cell fraction from a sample volume, comprising:

disrupting the sample volume to obtain a sample particle in a selected particle size range different than a sample volume particle size range prior to the disruption of the sample volume;

separating the selected cell fraction from the sample particle, including,

disposing the sample particle in a first section of a separation container;

applying a first centrifugal force to the sample particle in the first section of the separation container;

collecting the selected cell fraction in a second section of the separation container;

removing the selected cell fraction from the second section of the separation container.

2. The method of claim 1, further comprising:

spinning the separation container around a central axis to apply the centrifugal force to the sample particle;

moving the sample particle along an interior wall of the first section of the separation container while spinning the separation container; and

transferring the selected cell fraction into the second section of the separation container while spinning the separation container.

3. The method of claim 1, further comprising:

mixing the at least one of the sample volume or the sample particle with citrate, a biologically acceptable acid, or EDTA prior to separating the selected cell fraction.

4. The method of claim 1, wherein disrupting the sample volume to obtain a sample particle in a selected particle size range includes forming the sample particle to have an average particle size in a range of about 0.1 mm to about 5 mm.

5. The method of claim 1, wherein disrupting the sample volume to obtain a sample particle in a selected particle size range includes forming the sample particle to have an average particle size in a range of about 0.5 mm to about 2 mm.

6. The method of claim 1, wherein disrupting the sample volume to obtain a sample particle in a selected particle size range includes forming the sample particle to have an average particle size of about 1 mm.

7. The method of claim 1 further comprising:

disrupting the sample volume substantially only with a mechanical force applied to the sample volume to obtain the sample particle.

8. The method of claim 7, further comprising:

obtaining the sample volume from a source body; and

applying the selected cell fraction to the source body.

9. The method of claim 7, further comprising:

obtaining the sample volume from a volume of adipose tissue of a patient;

wherein disrupting the sample volume includes forming a plurality of sample particles.

10. The method of claim 1, further comprising:

prior to applying a centrifugal force, forming a seal between the first section of the separation container and the second section; and

removing the seal from between the first section of the separation container and the second section of the separation container upon application of an opening force due at least in part to the applied centrifugal force.

11. The method of claim 10, wherein applying the centrifugal force includes applying at least a first centrifugal force and a second opening centrifugal force, wherein the second opening centrifugal force includes the opening force;

wherein applying the second opening centrifugal force causes at least a portion of the selected cell fraction to exert a force on a lid that closes a passage between the first section to the second section to open the lid to allow the selected cell fraction to move from the first section to the second section allowing the collecting of the selected cell fraction.

12. The method of claim 10, further comprising:

flexing a portion of a lid to remove the seal from between the first section and the second section with the second opening centrifugal force.

13. The method of claim 10, further comprising:

moving a lid from a first lid position that is sealing the first section from the second section to a second lid position that is unsealing the first section from the second section.

14. The method of claim 1, further comprising:

collecting the selected cell fraction in a second section of the separation container includes moving the selected cell fraction to a sump; and

removing the selected cell fraction from the second section of the separation container includes applying a suction to the sump to remove the selected cell fraction.

15. A method of separating a selected cell fraction from a sample volume, comprising:

disrupting the sample volume to obtain a separable sample having a sample particle in a selected particle size range;

separating the selected cell fraction from the sample particle, including,

disposing the separable sample having the sample particle in a first section of a separation container having a first wall extending from a first bottom wall to an upper rim having an interior surface and an exterior surface;

applying a centrifugal force to the sample particle in the first section of the separation container to cause both (1) collecting the selected cell fraction in a collection section positioned near the upper rim and at least contacting the exterior surface of the first wall, the collection section having a second wall extending from a second bottom wall to the upper rim, and wherein the second bottom wall is nearer the upper rim than the first bottom wall and (2) moving a sealing section that seals a collection volume within the collection section when in a closed position to an open position by flexing a flexible section to allow the sealing section to move to the open position from the closed position; and

removing the selected cell fraction from the collection section of the separation container.

16. The method of claim 15, further comprising:

operating a drive motor to spin the separation container and the collection section simultaneously to apply the centrifugal force.

17. The method of claim 16, further comprising:

closing a housing to enclose all of the separation container, the collection section, the lid, the sealing section, the flexible section, and the drive motor.

18. The method of claim 15, further comprising:

placing a lid assembly extending over both the separation container and the collection section;

wherein in the closed position the sealing section selectively seals the collection volume from a container volume of the separation container;

wherein at a selected spin rate the applied centrifugal force the sealing section moves to the open position from the closed position;

applying a centrifugal force to the sample particle in the first section of the separation container to cause both (1) collecting the selected cell fraction in a collection section positioned near the upper rim and at least contacting the exterior surface of the first wall, the collection section having a second wall extending from a second bottom wall to the upper rim, and wherein the second bottom wall is nearer the upper rim than the first bottom wall and (2) moving a sealing section.

19. The method of claim 15, wherein disrupting the sample volume to obtain a separable sample having the sample particle in the selected particle size range includes forming the sample particle to have an average particle size in a range of about 0.1 mm to about 5 mm.

20. The method of claim 15, wherein disrupting the sample volume to obtain a separable sample having the sample particle in the selected particle size range includes forming the sample particle to have an average particle size in a range of about 0.5 mm to about 2 mm.

21. The method of claim 15, wherein disrupting the sample volume to obtain a separable sample having the sample particle in the selected particle size range includes moving the sample volume though a perforated member defining at least a through bore.

22. The method of claim 21, wherein moving the sample volume though a perforated member defining at least a through bore includes:

placing the sample volume in a sample containing volume having an end covered by the perforated member; and

moving a forcing member to force the sample volume through the perforated member to create the sample particle.

23. The method of claim 15, wherein removing the selected cell fraction from the collection section of the separation container includes:

drawing a vacuum within the collection section wherein a barrier wall defines a first end of the collection section on a first side of the barrier wall and a second end of the collection section on a second side of the barrier wall such that the barrier wall disrupts a complete annular well of the collection section;

wherein a vacuum is operable to be formed near the first end of the collection section to draw substantially all material out of the collection section through a port.

24. The method of claim 15, further comprising:

connecting a withdrawal device to a passage defined through a housing that encloses the separation container and the collection section.

25. A method of separating a selected cell fraction from a sample volume, comprising:

forming a separable volume having a sample particle in a selected particle size range from the sample volume;

disposing the formed sample particle in a first section of a separation container;

applying a first centrifugal force to the sample particle in the first section of the separation container while the first section is sealed relative to a second section of the separation container to cause at least a portion of the selected cell fraction to move within the first section of the separation container;

applying a second centrifugal force to the sample particle in the first section of the separation container to cause a sealing member to move from a sealed position to an unsealed position;

applying a third centrifugal force to the sample particle in the first section of the separation container after causing the sealing member to move to the unsealed position to allow at least the portion of the selected cell fraction to move into the second section of the separation container; and

removing at least a portion of at least the portion of the selected cell fraction from the second section of the separation container.

26. The method of claim 25, wherein applying the centrifugal force includes spinning the separation container around a central axis which causes at least the selected cell fraction to move along an interior wall of the first section of the separation container and transfer at least the portion of the selected cell fraction into the second section of the separation container.

27. The method of claim 25, further comprising:

prior to applying the first centrifugal force, forming a seal between the first section of the separation container and the second section of the separation container with the sealing member.

28. The method of claim 27, wherein applying the first centrifugal force and applying the second centrifugal force are the same force at a first time and a second time, respectively.

29. The method of claim 28, wherein applying the second centrifugal force causes the seal member to move to the unsealed position.

30. The method of claim 29, wherein applying the second centrifugal force includes causing at least a portion of the selected cell fraction to exert a force on the sealing member to allow the selected cell fraction to move from the first section to the second section.

31. The method of claim 25, wherein applying at least the second centrifugal force includes flexing a portion of the sealing member to remove the seal from between the first section and the second section.

32. The method of claim 25, further comprising:

moving the selected cell fraction to a sump; and

applying a suction to the sump to remove the selected cell fraction.

33. The method of claim 25, further comprising:

moving the selected cell fraction to a sump by ceasing the application of the first centrifugal force, ceasing the application of the second centrifugal force, and ceasing the application of the third centrifugal force.

34. The method of claim 25, further comprising:

mixing the at least one of the sample volume or the sample particle with citrate, a biologically acceptable acid, or EDTA prior to at least one of forming a separable volume, disposing the formed sample particle in a first section of a separation container, applying a first centrifugal force, applying a second centrifugal force, or applying a third centrifugal force.

35. The method of claim 25, wherein forming the separable volume having the sample particle in the selected particle size range includes forming the sample particle to have an average particle size in a range of about 0.5 mm to about 2 mm.

36. The method of claim 25, wherein forming the separable volume having the sample particle in the selected particle size range includes disrupting the sample volume with only a mechanical force applied to the sample volume to obtain the sample particle.

37. The method of claim 36, wherein disrupting the sample volume with only a mechanical force includes moving the sample volume though a perforated member defining at least a through bore.

38. The method of claim 37, wherein moving the sample volume though a perforated member defining at least a through bore includes:

placing the sample volume in a sample containing volume having an end covered by the perforated member; and

moving a forcing member to force the sample volume through the perforated member to create the sample particle.

39. The method of claim 36, further comprising:

obtaining the sample volume from a source body; and

applying the selected cell fraction to the source body.