International Journal of Cell Cloning, Vol 2, 9-19, Copyright © 1984 by AlphaMed Press
The detection of subclasses of granulosa cells with differing responsiveness to EGF, FGF, and gonadotropin preparations using an anchorage-independent clonal agar assay
I Bertoncello and TR Bradley
Subpopulations of granulosa cells of differing responsiveness to epidermal
growth factor (EGF), fibroblast growth factor (FGF), pituitary gonadotropin
preparations, and rat erythrocyte suspensions (RBC) have been detected
using an anchorage-independent clonal agar assay. All growth factor
preparations were capable of enhancing colony formation when added alone,
and elicited cloning efficiencies as high as 35% when added to the culture
system at optimal concentrations in a variety of combinations. The FGF
preparation was the single most effective stimulator of colony formation,
augmenting both colony numbers and colony size at concentrations as low as
50 ng/ml. However, unlike the other growth factors in this assay system, a
plateau in responsiveness could not be reached even at levels as high as 1
microgram/ml. NIH-FSH-P2 and NIAMMD-bLH-4 were considerably less potent
than other growth factors. Both preparations were inactive at
concentrations less than 1 microgram/ml and produced an optimal response at
10 micrograms/ml.
