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

Author(s): Fowke KR, Behnke J, Hanson C, Shea K, Cosentino LM

Publication: J Immunol Methods, 2000, Vol. 244, Page 139-44

PubMed ID: 11033026 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of storage at -30, -70, and -150 degrees C for 10 weeks on cell viability, levels of apoptosis, and the ability to be transformed by Epstein-Barr virus (EBV) of whole blood versus matched peripheral blood mononuclear cell (PBMC) specimens.

   Summary of Findings:

   A significant decrease in viability was detected in PBMC specimens stored at -30 degrees C compared to those stored at -70 or -150 degrees C (p<0.0001). Higher levels of spontaneous and activation-induced apoptosis were observed for both whole blood and PBMC specimens stored at -30 degrees C (p<0.001) than at -70 or -150 degrees C. The levels of spontaneous apoptosis in whole blood specimens at -70 or -150 degrees C were higher than those observed in matched PBMC specimens (p<0.005). EBV transformation of specimens stored at -30 degrees C was only possible in whole blood due to insufficient cell numbers from PBMC. In the whole blood specimens stored at -30 degrees C, transformation was only moderately successful in 1 of 4 specimens after 6 weeks. Specimens stored at lower temperatures were all able to be transformed in 2-4 weeks, however, the whole blood specimens appeared to initially lag behind the PBMC specimens with respect to transformation.

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Morphology	Fluorescent microscopy
   Morphology	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Peripheral blood mononuclear cells Whole blood
   Storage	Storage temperature	-30 degrees C -70 degrees C -150 degrees C
   Storage	Storage duration	10 d

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