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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
Author(s) -
Serke Stefan,
Säuberlich Sabine,
Huhn Dieter
Publication year - 1991
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.1991.tb00527.x
Subject(s) - haematopoiesis , flow cytometry , progenitor cell , myeloid , in vitro , cd15 , cell culture , biology , microbiology and biotechnology , immunology , cd34 , chemistry , stem cell , biochemistry , genetics
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 CD2 1 ‐positive cell‐depleted PBMC from normal subjects have been cultured in the presence of rhEPO, rhGM‐CSF or rhII‐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 myeloid cells with serum deprivation. Using dual‐colour flow‐cytometry, cell‐cycle analysis of CD13+ cells, comparing the effects of rhGM‐CSF to those of rhII‐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.