Two-day collection and pooling of peripheral blood stem cells with semiautomated density gradient cell separation

Wilbur J. Pan, Paul Haut, Marie Olszewski, Morris Kletzel

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Autologous and allogeneic PBSC collection and cryopreservation have been shown to be feasible in the pediatric population. This technique may be associated with complications, including volume overload and DMSO toxicity. To decrease these risks, we developed a technique by which PBSC are collected over a 2-day period and pooled prior to cryopreservation. PBSC are harvested on day 1 and stored with tissue culture medium on a rocker at ambient temperature. On day 2, a second PBSC harvest is performed, and the two harvests are pooled, separated on a Ficoll gradient in a semiautomatic closed system, and frozen with 10% DMSO after a soft spin for volume reduction. Cells from each day's harvest are tested for cell count and viability. A total of 36 collections in 26 patients were performed. This technique resulted in a 73% ± 0.0% reduction in volume (mean ± SEM) and an 88% ± 0.9% depletion of RBC. Mononuclear cell (MNC) count recovery was 88% ± 2.6%, and the MNC dose delivered to the patient was 3.1 ± 0.6 x 108 cells/kg. Cell viability was >98% before and after processing. Seventeen patients have been transplanted thus far, and all these patients engrafted with minimal toxicity. These data indicate that storing PBSC for up to 24 h after harvest does not decrease PBSC viability or delay engraftment.

Original languageEnglish (US)
Pages (from-to)561-564
Number of pages4
JournalJournal of Hematotherapy and Stem Cell Research
Volume8
Issue number5
DOIs
StatePublished - Oct 1999
Externally publishedYes

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Cell Separation
Cryopreservation
Dimethyl Sulfoxide
Cell Survival
Cell Count
Ficoll
Culture Media
Pediatrics
Temperature
Peripheral Blood Stem Cells
Population

ASJC Scopus subject areas

  • Hematology
  • Immunology

Cite this

Two-day collection and pooling of peripheral blood stem cells with semiautomated density gradient cell separation. / Pan, Wilbur J.; Haut, Paul; Olszewski, Marie; Kletzel, Morris.

In: Journal of Hematotherapy and Stem Cell Research, Vol. 8, No. 5, 10.1999, p. 561-564.

Research output: Contribution to journalArticle

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