NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

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

The purpose of this paper 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) on whole blood and matched peripheral blood mononuclear cell (PBMC) specimens.

Conclusion of Paper

Decreased cell viability and increased spontaneous and activation-induced apoptosis were observed when either whole blood or PBMC specimens were stored at -30 degrees C. In addition, higher levels of apoptosis were observed in whole blood specimens compared to PBMC specimens at the lower storage temperatures. EBV transformation was achievable in all specimens stored at -70 or -150 degrees C, but in only 1 of 4 specimens of whole blood stored at -30 degrees C.

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
    Preservative Types
    • Frozen
    Diagnoses:
    • Normal
    Platform:
    AnalyteTechnology Platform
    Morphology Fluorescent microscopy
    Morphology Light microscopy
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(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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