PMID: 6983974Nov 1, 1982Paper

Enrichment of hematopoietic progenitor cells (CFUC and BFUE) from human peripheral blood

Experimental Hematology
D WisniewskiB Clarkson

Abstract

Granulocytic (CFUC) and erythroid (BFUE) progenitor cells have been rapidly purified from human peripheral blood approximately 140 fold by combining centrifugation on a density cushion and immunoadherence cell separation methods. An initial light density (d less than 1.071 g/cm3) mononuclear cell fraction, enriched for progenitor cells, was obtained by centrifugation of whole blood on a modified Ficoll-Hypaque density cushion. Cultures of the light-density cells gave cloning efficiencies (defined as the percentage of total cells plated) of 0.008% and 0.015% for CFUC and BFUE respectively. Further purification was achieved by negative selection whereby selective populations of immunocompetent cells were removed. Thus, B cells and monocytes (as well as up to 50% high affinity Fc receptor bearing cells) were simultaneously depleted by immunoadherence to plastic petri dishes coated with rabbit anti-human IgG. Leu-3a positive (helper) and Leu-2a positive (suppressor) T cells were then simultaneously depleted by an indirect "panning" method, whereby the T cell subsets were coated with the corresponding murine monoclonal antibodies prior to their removal by immunoadherence to plastic petri dishes coated with goat anti-mouse IgG. The f...Continue Reading

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