Systems and methods for cryopreservation of cells

• US 8,222,027 B2
• Filed: 12/17/2008
• Issued: 07/17/2012
• Est. Priority Date: 06/20/2006
• Status: Active Grant

• Current Assignee: Sexton Biotechnologies, Inc. (Biolife Solutions, Inc.)
• Original Assignee: Cook General Biotechnolgy LLC (Cook Group Incorporated)
• Inventors: Woods, Erik John, Critser, John K.
• Primary Examiner(s): Beisner, William H
• Application Number:
  US12/337,237
• Publication Number:
  US 20090148934A1
• Time in Patent Office: 1,308 Days
• Field of Search: 435/297.1, 435/307.1, 435/297.5, 604/408, 604/411, 604/415, 624/579
• US Class Current: 435/307.1
• 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 that is 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. One such cryopreservation vessel is a flexible vial having a closed port at one adapted to be pierced by a needle to withdraw the liquid within, 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. In another embodiment, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Claims

1. A cryopreservation device comprising:

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

an auto-nucleating device disposed within said container, said auto-nucleating device including;

an elongated hollow tube sized for introduction through said inlet fitting;

an ice-nucleating composition disposed within said hollow tube; and

a seal at both ends of said tube, at least one end sealed with a membrane that is impermeable to the ice-nucleating composition but permeable to the liquid contained within the cryopreservation vessel;

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.

2. The cryopreservation device of claim 1, wherein said tube engaging fitting is a barbed fitting.

3. The cryopreservation device of claim 1, wherein said closure is a septum adapted to be pierced by a needle.

4. The cryopreservation device of claim 1, 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.

5. The cryopreservation device of claim 1, wherein said closure is formed by heat sealing said opposite end of said tube.

6. The cryopreservation device of claim 1, wherein said container is configured to be maintained at a pressure below atmospheric pressure.