SYSTEMS AND METHODS FOR CRYOPRESERVATION OF CELLS

• US 20130065301A1
• Filed: 07/16/2012
• Published: 03/14/2013
• Est. Priority Date: 06/20/2006
• Status: Active Grant

• Current Assignee: Sexton Biotechnologies, Inc. (Biolife Solutions, Inc.)
• Original Assignee: Cook General BioTechnology LLC (Cook Group Incorporated)
• Inventors: Woods, Erik John, Critser, John K.
• Granted Patent: US 8,936,905 B2
• Time in Patent Office: Days
• US Class Current: 435/374
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0231 Chemically defined matrices, e.g. alginate gels, for immobilising, holding or storing cells, tissue or organs for preservation purposes; Chemically altering or fixing cells, tissue or organs, e.g. by cross-linking, for preservation purposes
• CPC Class Codes: A01N 1/0268 Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen

Abstract

An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. The cryopreservation vessel can be a flexible vial having a closed port adapted to be pierced by a needle, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. Alternatively, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Claims

1-13. -13. (canceled)

14. A cryopreservation device comprising:

a container for receiving and storing a liquid sample, said container having an inlet fitting opening into said container;

an adaptor mounted to said fitting, said adaptor having a first tubular branch and a second tubular branch, said second tubular branch terminating in a tube engaging fitting;

a septum closing said first tubular branch, said septum adapted to be pierced by a needle;

a tube engaged at one end to said tube engaging fitting on said second tubular branch; and

a closure at the opposite end of said tube.

15. The cryopreservation device of claim 14, wherein said fitting is a barbed fitting.

16. The cryopreservation device of claim 14, wherein said closure is a septum adapted to be pierced by a needle.

17. The cryopreservation device of claim 14, wherein said tube is a flexible tube having a length so that said opposite end of said tube is above said septum closing said first branch when said container is upright.

18. The cryopreservation device of claim 14, wherein said closure is formed by heat sealing said opposite end of said tube.

19. The cryopreservation device of claim 14 further comprising an auto-nucleating device disposed therein.

20. The cryopreservation device of claim 14, wherein said container is maintained at a pressure below atmospheric pressure.

21. A cryopreservation device comprising:

a container for receiving and storing a liquid sample, said container having an upper opening and a lower opening;

a cap in sealed engagement with said container covering said upper opening, said cap including a first tubular branch and a second tubular branch communicating with said upper opening of said container; and

an outlet septum closing said lower opening of said container, said outlet septum adapted to be pierced by a needle to withdraw the liquid sample from said container.

22. The cryopreservation device of claim 21, further comprising an inlet septum in said first tubular branch, said inlet septum adapted to be pierced by a needle to inject the liquid sample into said container.

23. The cryopreservation device of claim 21, further comprising a micro-filter disposed within said second branch, said micro-filter being permeable to gas.

24. The cryopreservation device of claim 23, wherein said micro-filter is impermeable to the liquid sample being stored in said container.

25. The cryopreservation device of claim 21, wherein said first and second branch are heat sealed when the liquid sample is disposed within said container.

26. The cryopreservation device of claim 21, further comprising a removable cover in sealed engagement with said container surrounding said outlet septum.

27. A method, comprising:

cryogenically storing a cryopreservation container having a chamber defined between a first opening and a second opening and a cellular composition received in the chamber, the cryopreservation container further including a first sealed tubular branch communicating with said first opening, a second sealed tubular branch communicating with said first opening, and a septum closing said second opening of said container, said septum adapted to be pierced by a needle.

28. The method of claim 27, also comprising:

after said cryogenically storing, subjecting the cryopreservation container to conditions sufficient to thaw the cellular composition.

29. The method of claim 28, also comprising:

piercing the septum with a needle; and

withdrawing an amount of the cellular composition through the needle.

30. The method of claim 27, wherein the first and second sealed tubular branches each include a heat-sealable tube material having a heat-sealed region of the tube material.

31. A method, comprising:

passing a liquid cellular suspension through a first tubular member and into a chamber defined by a cryopreservation container;

during said passing, venting gas from within the chamber out a second tubular member in fluid communication with the chamber.

32. The method of claim 31, wherein the cellular suspension includes a cryopreservative.

33. The method of claim 31, wherein the cryopreservation container also includes a needle septum communicating with said chamber.

34. The method of claim 33, wherein the needle septum is positioned in the second tubular member.

35. The method of claim 33, wherein the chamber is defined between a first opening and a second opening of the container, wherein the first and second tubular members communicate with the first opening, and wherein the needle septum seals the second opening.

36. The method of claim 31, also comprising cryogenically storing the cryopreservation container with the cellular suspension received in the chamber and the first and second tubular members in a sealed condition.