Methodological studies on growth of mouse granulocyte/macrophage colonies in methylcellulose-containing media.
We studied the influence of various physicochemical parameters on colony development and total cell numbers in 7-day methylcellulose cultures of mouse bone marrow cells. Colony growth was markedly retarded by an increase of PO2 from approximately 6.7 kPa towards that in ambient air and by a decrease of incubator temperature from 37 degrees C to 33 degrees C. Medium osmolality above approximately 340 mosm/kg inhibited formation of granulocytes (in cultures containing growth factors from pokeweed mitogen-stimulated spleen cells), but not macrophages (L cell-produced growth factors). At most, colony growth was affected slightly by moderate changes in pH (7.17-7.47) or concentration of methylcellulose (0.77-1.02%), or by the presence of 2-mercaptoethanol (50 mumol/1), Hepes buffer (25 mmol/1), or erythropoietin (0.1-1 units/ml). The culture trays could be stored for one day at 4 degrees C in the incubation boxes before colonies or cells were counted.
D000818:Animals; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D003470:Culture Media; D005260:Female; D006098:Granulocytes / Q000502:physiology*; D008264:Macrophages / Q000502:physiology*; D008747:Methylcellulose; D051379:Mice
D003470:Culture Media; D008747:Methylcellulose