Viability and recovery of peripheral blood mononuclear cells cryopreserved for up to 12 years in a multicenter study.

Author(s): Kleeberger CA, Lyles RH, Margolick JB, Rinaldo CR, Phair JP, Giorgi JV

Publication: Clin Diagn Lab Immunol, 1999, Vol. 6, Page 14-9

PubMed ID: 9874657 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 frozen storage of up to 12 years on the recovery and viability of PBMC collected from HIV-seropositive and HIV-seronegative donors at 4 different centers.

   Summary of Findings:

   Across all of the centers, an average recovery of 77.5% with a median viability of 92% was observed. The percent recovery and viability were dependent on the center at which freezing and thawing occurred, but all centers had at least a median viability of 90%. All centers showed a trend toward decreasing viability with storage and an average decline of 2.9% over the 12 year period, but this was not significant for any of the centers. No difference in viability was observed based on HIV-serostatus. The median number of CD4+ lymphocytes showed a strong positive correlation with storage in two centers, but no relationship at the other two indicating that immunosuppression did not decrease PBMC viability.

   Biospecimens

   • Cell - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • AIDS/HIV-related
   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   Cell count/volume	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0-12 years
   Storage	Storage temperature	-135 degrees C or below
   Biospecimen Aliquots and Components	Blood and blood products	Peripheral blood mononuclear cells
   Preaquisition	Diagnosis/ patient condition	HIV-seronegative HIV-seropositive

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2. Study Purpose

   The purpose of this study was to determine if freezing PBMC in isopropanol insulated containers versus controlled rate freezers alters recovery and viability after storage.

   Summary of Findings:

   Specimens which had been frozen in controlled rate freezers only had a 0.01% loss of viable cells per year of storage while those stored in isopropanol insulated containers showed an average loss of 2.23% per year. When this study was expanded to other centers the effect was not significant; however, specimens frozen in alcohol filled containers consistently had a lower percent of viable PBMC when viability cut-offs were used.

   Biospecimens

   • Cell - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • AIDS/HIV-related
   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   Cell count/volume	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Peripheral blood mononuclear cells
   Storage	Storage duration	8-12 years 0-8 years 0-12 years
   Storage	Storage temperature	-135 degrees C or below
   Biospecimen Preservation	Cooling or freezing method/ rate	Isopropanol insulated containers Controlled rate freezers

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