Protective effect of L-glutamine against freeze-thaw damage in mammalian cells

L-Glutamine at 18 mM protects mammalian cells against freeze-thaw (FT) damage by a factor of about 6, depending on FT conditions, in balanced salt solutions. While not nearly as effective a cryoprotectant as dimethyl sulfoxide (DMSO) or propylene glycol (PG), the mechanism of protection by glutamine appears to be independent from that of DMSO or PG; thus, 18 mM glutamine is effective at reducing FT damage in combination with these agents. These combinations allow lower concentrations of the more toxic agents DMSO and PG to be used in FT medium. There is no pre-FT or post-FT effect of glutamine when cells are exposed to a FT cycle in balanced salt solutions. Hence, protection is due to its presence during the FT-cycle. The presence of 2 mM L-glutamine in Eagle's basal medium is sufficient to account for cryoprotection by this medium.