Defined, serum-free culture conditions for the GM-micro-clonogenic assay using agar-capillaries.
Culture conditions were determined and optimised for the serum-free growth of granulocyte-macrophage (GM) colonies, derived from mouse bone marrow cells, in agar-containing glass capillaries. The standard medium is DMEM/F-12 (1:1) mixture containing per ml: 10 mg BSA, 35 micrograms transferrin, 20 micrograms soybean lipids and 5 micrograms insulin. In contrast to previous attempts by others, GM-colony yield in serum-free medium was found to be equal to that in serum-containing medium (around 25 colonies/capillary) and to correlate satisfactorily with the cell density at seeding.--The role of polyamine oxidases in cell-proliferation experiments could be studied by this microclonogenic assay without interference from any type of serum.
Institut für Pharmazie der Freien Universität, Berlin.
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Maurer
D000362:Agar; D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D002452:Cell Count; D002478:Cells, Cultured; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods*; D019856:Ethanolamine; D004983:Ethanolamines / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology* / Q000187:drug effects; D008264:Macrophages / Q000166:cytology* / Q000187:drug effects; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D014168:Transferrin / Q000494:pharmacology
D003470:Culture Media; D004983:Ethanolamines; D014168:Transferrin; D012710:Serum Albumin, Bovine; D013096:Spermine; D019856:Ethanolamine; D000362:Agar