International Journal of Cell Cloning, Vol 2, 113-125, Copyright © 1984 by AlphaMed Press
Methodological studies on growth of mouse granulocyte/macrophage colonies in methylcellulose-containing media
HB Benestad, IB Hersleth and I Strom-Gundersen
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.
