In vitro assessment of cell viability in human bone marrow preserved at –79 C

¹ Departments of Radiotherapy and Haematology, Westminster Hospital and Medical School, London, England

Human bone marrow was stored at –79 C for 1 hr with glycerol, with dimethyl sulfoxide, or without any protective agent, both after rapid cooling and after slow cooling. Each sample was subjected to four viability tests, and observations were also made on samples stored for 1 hr at room temperature. By comparison with the established effects of these preservation procedures on the radiation recovery factor, it is concluded that: 1) the dye exclusion test using trypan blue is a poor indicator of viability after freezing; 2) the measurement of deoxyribonucleic acid synthesis gives results which correlate well with in vivo assays; 3) the detection of motility is the only in vitro test to provide unequivocal evidence of life, but is not susceptible to quantitative measurement: slow cooling in the presence of glycerol or dimethyl sulfoxide was the most successful preservation procedure used; 4) supravital staining with acridine orange, although it is a purely empirical test, gives a good correlation with the results of in vivo assay and is simple, convenient, and rapid to perform.

Submitted on May 20, 1963