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.
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.
D000362:Agar; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002999:Clone Cells; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000166:cytology*
D004815:Epidermal Growth Factor; D000362:Agar