Abstract
To facilitate the direct study of progenitor cell biology, we have developed a simple and efficient procedure based upon negative selection by panning to purify large numbers of committed erythroid and myeloid progenitors from human fetal liver. The nonadherent, panned cells constitute a highly enriched population of progenitor cells, containing 30.4 +/- 13.1% erythrocyte burst forming units (BFU-E), 5.5 +/- 1.9% granulocyte-macrophage colony forming units (CFU-GM), and 1.4 +/- 0.7% granulocyte-erythroid-macrophage-megakaryocyte colony forming units (CFU-GEMM) as assayed in methylcellulose cultures. These cells are morphologically immature blasts with prominent Golgi. This preparative method recovers 60-100% of the committed progenitors detectable in unfractionated fetal liver and yields 2-30 X 10(6) progenitors from each fetal liver sample, and thus provides sufficient numbers of enriched progenitors to allow direct biochemical and immunologic manipulation. Using this technique, a purified recombinant protein previously thought to have only granulocyte-macrophage colony stimulating activity (GM-CSA) is shown to have both burst promoting activity and multipotential colony stimulating activity. Progenitor purification by panning thus appears to be a simple, efficient method that should facilitate the direct study of committed hematopoietic progenitors and their differentiation.
MeSH Terms
Cell Count
Cell Separation/methods
Colony-Forming Units Assay
Colony-Stimulating Factors/physiology
Erythrocytes/cytology
Erythropoiesis
Fetus
Hematopoiesis
Hematopoietic Stem Cells/cytology
Humans
Liver/cytology
Recombinant Proteins/physiology
Chemicals
Colony-Stimulating Factors
Recombinant Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Emerson S G
Sieff C A
Wang E A
Wong G G
Clark S C
Nathan D G
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