Optimization of storage and shipment of cryopreserved peripheral blood mononuclear cells from HIV-infected and uninfected individuals for ELISPOT assays.

Author(s): Weinberg A, Song LY, Wilkening CL, Fenton T, Hural J, Louzao R, Ferrari G, Etter PE, Berrong M, Canniff JD, Carter D, Defawe OD, Garcia A, Garrelts TL, Gelman R, Lambrecht LK, Pahwa S, Pilakka-Kanthikeel S, Shugarts DL, Tustin NB

Publication: J Immunol Methods, 2010, Vol. 363, Page 42-50

PubMed ID: 20888337 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 duration and shipping temperature on viability and proliferation of PBMC from HIV positive and negative individuals. Cell were cryopreserved within 8 h of blood collection and stored in liquid nitrogen or -70 degrees C. Replicate specimens stored in liquid nitrogen were shipped on dry ice after 6 weeks or in liquid nitrogen after 7 weeks, while replicate specimens stored at -70 degrees C were shipped on dry ice at weekly intervals.

   Summary of Findings:

   The mean viability and recovery were each 5% lower (p<0.01 and p=0.04, respectively) when specimens stored in liquid nitrogen were shipped on dry ice instead of liquid nitrogen. Specimens stored at -70 degrees C for 3 weeks and shipped on dry ice had significantly lower mean viability (75%) and recovery (55%) than specimens stored in liquid nitrogen and shipped either in liquid nitrogen (89% and 70%, respectively) or on dry ice (84% and 65%, respectively). Generally, HIV status did not affect PBMC viability or recovery, but the viability of PBMC from HIV infected individuals stored in liquid nitrogen and shipped on dry ice was reduced compared to that of specimens from uninfected individuals with the same storage and shipping conditions. PBMC viability and recovery decreased linearly with increasing storage duration at -70 degrees C in specimens from HIV positive and negative individuals, but the decline was faster in specimens from HIV positive individuals. ELISpot results were not different between specimens stored in liquid nitrogen and shipped in liquid nitrogen or on dry ice and those stored at -70 degrees C for 3 weeks and shipped on dry ice. In response to cytomegalovirus (CMV) pp65, CEF, and candida albicans, ELISpot values decreased with storage at -70 degrees C, but only the declines in response to CMV pp65 (p=0.03) and candida albicans (p=0.007) reached significance. 85% of specimens stored in liquid nitrogen and shipped on dry ice, 81% of specimens stored for 3 weeks at -70 degrees C, 75% of specimens stored for 6 weeks at -70 degrees C, 64% of specimens stored for 9 weeks at -70 degrees C, and 60% of specimens stored at -70 degrees C for 12 weeks had ELISpot results in response to CMV pp65 within 2 fold of those obtained using specimens stored and shipped on liquid nitrogen.

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • AIDS/HIV-related
   • Not specified

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Light microscopy
   Cell count/volume	ELISpot

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Between site transportation method	Mailed
   Storage	Specimen transport duration/condition	Dry ice Liquid nitrogen
   Storage	Storage duration	3 weeks 6 weeks 7 weeks 9 weeks 12 weeks
   Preaquisition	Diagnosis/ patient condition	HIV positive HIV negative
   Storage	Storage temperature	Dry ice Liquid nitrogen -70 degrees C

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