Stem Cells, Vol 12, 494-505, Copyright © 1994 by AlphaMed Press

ORIGINAL ARTICLES

Enrichment and characterization of peripheral blood-derived megakaryocyte progenitors that mature in short-term liquid culture

JL Nichol, AC Hornkohl, ES Choi, MM Hokom, I Ponting, FW Schuening and P Hunt
Amgen, Inc., Thousand Oaks, California 91320.

A population of peripheral blood-derived cells that mature into megakaryocytes within four to eight days of liquid culture is described. This population was enriched from normal leukapheresis units by counterflow centrifugal elutriation and CD34 selection. The cells were incubated in suspension with known megakaryocyte growth or maturation factors. Megakaryocytes were identified within the cultures with antibodies to platelet-glycoproteins (Ib and IIb) and cytologically classified as stage I-IV cells. Plasma from aplastic dogs (APK9) or human recombinant interleukin 3 (IL-3) were the only culture additives which reproducibly resulted in megakaryocyte development. The activity present in APK9 was relatively megakaryocyte-specific while IL- 3 was not. The phenotype of the short-term megakaryocyte progenitor cell population was determined by FACS and found to be CD34bright but not CD34dull or CD34-. The population was further characterized as CD34+/CD38+ and CD34+/HLA-DR+. Both CD34+/CD41- and CD34+/CD41+ populations contained megakaryocyte progenitor cells, although megakaryocytes that developed from the latter population were fewer in number. In summary, we have developed an enrichment protocol that, when coupled with a liquid culture assay system, enabled us to characterize short-term megakaryocyte progenitors from peripheral blood. These cells may be important for early platelet recovery in transplantation settings.

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