A liquid culture method for the in vitro growth of hemopoietic progenitor cells from normal human adult peripheral blood allowing for analysis by multiparameter flow-cytometry.
A liquid culture method has been developed allowing for the in vitro growth of peripheral blood-derived hemopoietic progenitor cells of myeloid, erythroid, monocytic and megakaryocytic lineages. Adherent cell- and CD21-positive cell-depleted PBMC from normal subjects have been cultured in the presence of rhEPO, rhGM-CSF or rhIl-3. Culturing cells in liquid cultures and in plasma clots, a similar dose-response was observed for granulocytic cells/liquid culture and granulocytic colonies/plasma clot with rhGM-CSF, and also for erythroid cells/liquid culture and erythroid colonies/plasma clot with rhEPO. Comparing serum- liquid cultures to serum+ liquid cultures, the ratio of CD13+ cells to CD15+ cells was higher in serum- cultures, indicating a maturation arrest of myecloid cells with serum deprivation. Using dual-colour flow-cytometry, cell-cycle analysis of CD13+ cells, comparing the effects of rhGM-CSF to those of rhIl-3, have been performed. The liquid culture method promises to be a useful tool for the study of in vitro differentiation and proliferation of hemopoietic progenitor cells.
Serke
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Universitätsklinikum Rudolf Virchow-Charlottenburg, Freie Universität Berlin, Federal Republic of Germany.
Säuberlich
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Huhn
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D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D000943:Antigens, Differentiation / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D018826:CD13 Antigens; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D015672:Erythroid Precursor Cells / Q000166:cytology; D004921:Erythropoietin / Q000494:pharmacology; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D016256:Lewis X Antigen; D018950:Lipopolysaccharide Receptors; D008533:Megakaryocytes / Q000166:cytology; D009000:Monocytes / Q000166:cytology
D015703:Antigens, CD; D000943:Antigens, Differentiation; D015214:Antigens, Differentiation, Myelomonocytic; D007377:Interleukin-3; D016256:Lewis X Antigen; D018950:Lipopolysaccharide Receptors; D004921:Erythropoietin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D018826:CD13 Antigens