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J Immunol Methods. 2000 Oct 20;244(1-2):139-44.

Apoptosis: a method for evaluating the cryopreservation of whole blood and peripheral blood mononuclear cells.

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Department of Cell Biology, BBI-Biotech Research Laboratories, 217 Perry Parkway, Gaithersburg, MD 20877, USA.


We sought to compare the effect of cryopreservation and storage at -30 degrees C, -70 degrees C and -150 degrees C of human whole blood versus matched peripheral blood mononuclear cell (PBMC) samples using apoptosis as an indicator of cell fitness. Following 10 weeks of storage the samples were thawed and assessed for viability (trypan blue exclusion), levels of apoptosis (using the nuclear stain bis-benzimide) and cell function (ability to be transformed by Epstein-Barr virus, EBV). When comparing storage temperatures, the levels of apoptosis in whole blood and PBMC samples stored at -30 degrees C were significantly higher than the values for samples stored at -70 degrees C or -150 degrees C (P<0.004). Whole blood samples stored at -150 degrees C had significantly less apoptosis than those stored at -70 degrees C (P<0.03). A comparison of the cell preparations showed that at all three storage temperatures there was significant sample deterioration (viability, apoptosis, and function) in whole blood relative to PBMC samples. This study indicates that careful consideration should be given to storage conditions and that apoptosis can be used as a sensitive measure of cell fitness following cryopreservation.

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