APPENDIX 3G Cryopreservation and Thawing of Cells

  1. Wayne M. Yokoyama1,
  2. Maria L. Thompson2,
  3. Rolf O. Ehrhardt2

Published Online: 1 NOV 2012

DOI: 10.1002/0471142735.ima03gs99

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Yokoyama, W. M., Thompson, M. L. and Ehrhardt, R. O. 2012. Cryopreservation and Thawing of Cells. Current Protocols in Immunology. 99:3G:A.3G.1–A.3G.5.

Author Information

  1. 1

    University of California School of Medicine, San Francisco, California

  2. 2

    BioCision LLC, Larkspur, California

Publication History

  1. Published Online: 1 NOV 2012
  2. Published Print: NOV 2012


Successful cryopreservation of cells requires not only that the cells be handled in a proper fashion for harvesting with equipment in place to ensure consistency, reproducibility, and sterility, but also that a correct choice and amount of cryoprotective agent is added. In general, a controlled freezing rate of 1°C/min is necessary to retain optimal viability of the recovered cells. There are many variations of cell freezing methods in use, including costly electronically regulated control rate freezers, unstandardized, passive isopropyl alcohol freezing containers, and crude rudimentary devices constructed from Styrofoam boxes or paper insulation. However, for the freezing and recovery of cell lines, primary cells, and stem cell cultures, the protocol described in this unit is simple, reproducible, and successful. Not only does it eliminate the need for isopropanol, as well as the costs and hazards associated with its use and disposal, but it provides a uniform method with improved cell viability and recovery. Curr. Protoc. Immunol. 99:A.3G.1-A.3G.5. © 2012 by John Wiley & Sons, Inc.


  • cell culture mammalian;
  • cell biology;
  • cell isolation and culture