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Human mixed cell colonies: unicellular or multicellular origin–analysis by G‐6‐PD
Author(s) -
Powell Jerry S.,
Fialkow Philip J.,
Adamson John W.
Publication year - 1984
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1984.tb02868.x
Subject(s) - multipotent stem cell , haematopoiesis , biology , in vitro , stem cell , microbiology and biotechnology , cell , enumeration , cell culture , plating efficiency , enzyme , colony forming unit , locus (genetics) , bone marrow , cell type , cell counting , flow cytometry , immunology , biochemistry , cell cycle , genetics , progenitor cell , gene , mathematics , combinatorics , bacteria
S ummary. Marrow and peripheral blood cells from normal women heterozygous (Gd B /Gd A ) at the X‐chromosome‐linked glucose‐6‐phosphate dehydrogenase (G‐6‐PD) locus were cultured at cell concentrations ranging from 2 × 10 4 /ml to 4 × 10 5 /ml to test formally the plating conditions necessary for reliable enumeration of multipotent stem cells (CFU‐mix). The culture system was rigorously tested by plating cells obtained after velocity sedimentation and the G‐6‐PD enzyme type of individual colonies was determined. At cell concentrations <7.5 × 10 4 /ml for marrow and < 1.25 × 10 5 /ml for peripheral blood, mixed‐cell colonies had either type A or type B enzyme, but not both. At higher cell concentrations, significant numbers of colonies showed both enzyme types and therefore arose from more than one cell. These studies demonstrate that enumeration of CFU‐mix by in vitro colony assay is accurate only at low cell concentrations. Studies of haematopoietic differentiation relying on in vitro colony assays of multipotent stem cells must be carefully analysed in light of these data.