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Clinical and Diagnostic Laboratory Immunology, January 1999, p. 14-19, Vol. 6, No. 1
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Viability and Recovery of Peripheral Blood Mononuclear Cells Cryopreserved for up to 12 Years in a Multicenter Study

Cynthia A. Kleeberger,1,* Robert H. Lyles,1 Joseph B. Margolick,2 Charles R. Rinaldo,3 John P. Phair,4 and Janis V. Giorgi5

Department of Epidemiology1 and Department of Molecular Microbiology and Immunology,2 Johns Hopkins School of Public Health, Baltimore, Maryland; Departments of Pathology and Microbiology, University of Pittsburgh Graduate School of Public Health, Pittsburgh, Pennsylvania3; Department of Medicine, Northwestern University School of Medicine and the Howard Brown Health Center, Chicago, Illinois4; and Department of Medicine and Jonsson Comprehensive Cancer Center, University of California School of Medicine, Los Angeles, California5

Received 22 June 1998/Returned for modification 11 August 1998/Accepted 12 October 1998

The Multicenter AIDS Cohort Study (MACS), an ongoing prospective study of the natural history of human immunodeficiency virus (HIV), has stored biologic specimens, including peripheral blood mononuclear cells (PBMC), from 5,622 participants for up to 12 years. The purpose of the present analysis was to evaluate the quality of the PBMC in the MACS repository in order to test the validity and feasibility of nested retrospective studies and to guide the planning of future repositories. PBMC were collected from MACS participants at four centers at 6-month intervals from 1984 to 1995, cryopreserved, and transported to a central repository for storage. A total of 596 of these specimens were subsequently tested for viability and used to evaluate cell function, to conduct immunophenotype analysis, or to isolate HIV. Simple linear regression models were applied to evaluate trends in recovery and viability over time and by center. Results indicated that from a nominal 107 cells cryopreserved per vial at all four centers, the median number of viable cells recovered was at least 5 × 106 (50% of the number stored) and the median viability was at least 90%. Results suggested that cryopreserved cells can be stored for at least 12 years with no general tendency toward cell loss over time. Furthermore, there were no statistically significant changes in the percent cell viability according to the length of time frozen, regardless of HIV serostatus or the level of CD4+ lymphocytes. Storing 107 PBMC per vial yields sufficient viable cells for phenotypic and/or functional analysis. Results from the MACS provide the basis for the planning of future repositories for use by investigators with similar research goals.

* Corresponding author. Mailing address: Johns Hopkins School of Public Health, Department of Epidemiology, 615 N. Wolfe St., Baltimore, MD 21205. Phone: (410) 955-4320. Fax: (410) 955-7587. E-mail: ckleeber{at}jhsph.edu.

Clinical and Diagnostic Laboratory Immunology, January 1999, p. 14-19, Vol. 6, No. 1
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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