International Journal of Cell Cloning, Vol 5, 142-148, Copyright © 1987 by AlphaMed Press
Anchorage-independence as an index of proliferative potential and maturational age: a comparison of the growth of normal, primary explanted bovine granulosa cells in semisolid agar and in liquid culture
RK Bartholomeusz and I Bertoncello
When seeded at low-density, normal primary explanted granulosa cells will
grow to form clones of functionally differentiated cells in both semisolid
agar and in liquid culture. The anchorage-independent clonogenic granulosa
cell differs from the anchorage-dependent granulosa cells detected in
clonal liquid culture in a number of properties. Basal cloning efficiency
in liquid culture is up to 50-fold higher than in agar culture. In serum
supplemented medium (20% fetal calf serum) cloning efficiency in liquid
culture is unaltered in the presence of added epidermal growth factor
(EGF), whereas, agar cloning efficiency is augmented six-fold when cells
are incubated under identical conditions. Cells derived from primary
anchorage-independent clones, when dispersed and replated, will generate
secondary anchorage- independent clones and anchorage-dependent liquid
clones. On the other hand, although cells derived from parallel primary
anchorage-dependent clones will also generate secondary anchorage-dependent
clones, generation of secondary anchorage-independent clones is not
detectable. These findings suggest that the anchorage-independent clonal
agar assay may be detecting a developmentally earlier granulosa cell
subpopulation than is detectable in the liquid culture assay.
