Device and method for controlled freezing of cell cultures

• US 4,455,842 A
• Filed: 09/27/1982
• Issued: 06/26/1984
• Est. Priority Date: 07/15/1981
• Status: Expired due to Fees

• Current Assignee: Aquila Biopharmaceuticals, Inc. (Agenus Inc.)
• Original Assignee: Biotech Research Laboratories Inc.
• Inventors: Granlund, David J.
• Primary Examiner(s): Capossela, Ronald C.
• Application Number:
  US06/423,744
• Time in Patent Office: 638 Days
• Field of Search: 62/78, 62/64, 62/457, 62/466
• US Class Current: 62/64
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0252 Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
• CPC Class Codes: A01N 1/0263 Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
• CPC Class Codes: C12N 1/04 Preserving or maintaining viable microorganisms immobilised microorganisms C12N11/00
• CPC Class Codes: F25D 3/00 Devices using other cold materials; Devices using cold-storage bodies
• CPC Class Codes: F25D 9/00 Devices not associated with refrigerating machinery and not covered by groups F25D1/00 - F25D7/00; Combinations of devices covered by two or more of the groups F25D1/00 - F25D7/00

Abstract

An enclosed device and method for controlled freezing of cell cultures, especially eucaryotic cell cultures, contained in cell culture vials by means of a conventional freezer includes a container and top, a supporting grid and supporting grid base within the container for supporting the culture vials and a compatible liquid refrigerant in a volume of at least fifteen times the culture volume and at a level to totally immerse all of the cultures to be frozen. Compatible liquid refrigerants include the straight chain aliphatic alcohols, preferrably ethanol and methanol.

Claims

1. A method of freezing cell cultures contained in one or more cell culture vials which comprises the steps of:

(a) at least partially filling a container with a prescribed volume of liquid refrigerant;

(b) supporting the culture vials so that the cell culture contained therein can be submersed in the liquid refrigerant;

(c) submersing at least the portion to the vial containing the cell culture into the liquid refrigerant so that the liquid refrigerant surrounds the vial portion submersed and the cell culture is below the refrigerant liquid level;

(d) said prescribed volume of liquid refrigerant being more than 15 times the total volume of cell culture to be frozen; and

(e) placing the container with culture vials submersed in the liquid refrigerant in a conventional freezer at a temperature below about -70°

C.

2. A method of controlling the freezing of cell cultures contained in one or more cylindrical cell culture vials which comprises supporting the culture vials in an upright position by means of a mechanical grid support so that the cell culture is in the lower portion of the cylindrical vial, submersing at least the lower portion of the cylindrical vial containing the cell culture in a volume of liquid refrigerant, the volume of liquid refrigerant being more than fifteen times the total volume of cell culture to be frozen, and placing the submersed culture vials in a conventional freezer at a temperature below about -70°

C.

3. The method of claim 2 wherein said liquid refrigerant is a straight chain aliphatic alcohol.

4. The method of claim 3 wherein said liquid refrigerant is selected from the group consisting of methanol and ethanol.

5. The method of claim 2 wherein the freezing is controlled to reduce the temperature of the cell culture at the rate of 1°

C. per minute plus or minus 20%.

6. The method of claim 5 wherein said cell cultures consist essentially of eucaryotic cell cultures.