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

Eur J Haematol. 1991 Feb;46(2):85-92. doi: 10.1111/j.1600-0609.1991.tb00527.x.

Abstract

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.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Antigens, CD / analysis
  • Antigens, Differentiation / analysis
  • Antigens, Differentiation, Myelomonocytic / analysis
  • CD13 Antigens
  • Cell Differentiation
  • Cell Division
  • Cells, Cultured
  • Erythroid Precursor Cells / cytology
  • Erythropoietin / pharmacology
  • Flow Cytometry*
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Granulocytes / cytology
  • Hematopoietic Stem Cells / cytology*
  • Humans
  • Interleukin-3 / pharmacology
  • Lewis X Antigen
  • Lipopolysaccharide Receptors
  • Megakaryocytes / cytology
  • Monocytes / cytology

Substances

  • Antigens, CD
  • Antigens, Differentiation
  • Antigens, Differentiation, Myelomonocytic
  • Interleukin-3
  • Lewis X Antigen
  • Lipopolysaccharide Receptors
  • Erythropoietin
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • CD13 Antigens