Standardized Cryopreservation of Human Primary Cells

Thomas V. Ramos, Aby J. Mathew, Maria L. Thompson, Rolf O. Ehrhardt

HemaCare Corporation, Van Nuys, California,  BioLife Solutions, Bothell, Washington
BioCision, Larkspur, California

Curr. Protoc. Cell Biol. 64:A.3I.1‐A.3I.8.


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Cryopreservation is the use of low temperatures to preserve structurally intact living cells. The cells that survive the thermodynamic journey from the 37°C incubator to the −196°C liquid nitrogen storage tank are free from the influences of time. Thus, cryopreservation is a critical component of cell culture and cell manufacturing protocols. Successful cryopreservation of human cells requires that the cells be derived from patient samples that are collected in a standardized manner, and carefully handled from blood draw through cell isolation. Furthermore, proper equipment must be in place to ensure consistency, reproducibility, and sterility. In addition, the correct choice and amount of cryoprotectant agent must be added at the correct temperature, and a controlled rate of freezing (most commonly 1°C/min) must be applied prior to a standardized method of cryogenic storage. This appendix describes how human primary cells can be frozen for long‐term storage and thawed for growth in a tissue culture vessel.


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