Factors affecting survival of mouse bone marrow cells during freezing and thawing in glycerol

¹ Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee

The effects of freezing rate, thawing rate and repeated freezing and thawing on the survival of mouse bone marrow cells frozen in various concentrations of glycerol were studied. The ability of thawed cells to promote recovery in lethally irradiated (900 r) mice was used as a measure of their viability. The experiments show that for best survival the cells should be ‘slow’ frozen in an additive solution, such as 15% glycerol. They should be ‘fast’ thawed. Repeated freezings and thawings lead to both a loss of cells and a more rapid loss of their ability to promote recovery. It is unnecessary to remove the glycerol from cells if they are to be used soon after thawing.