Estimation of viability of bovine granulosa cells.
Granulosa cells harvested from non-atretic, antral follicles of cow (and pig and sheep) ovaries were incubated over glass cover slips in medium containing 20% (v/v) donor calf serum. Cell attachment to the cover slips was rapid, being in most cases complete within 3 h at 37 degrees C. There was little further change over the next 20 h. The number of bovine granulosa cells which attached to a cover slip was proportional to the volume of cell suspension added to the medium; and the amount of oestradiol secretion by attached cells in a testosterone-enriched medium increased in parallel with their number. Granulosa cells which did not attach within 3 h produced little oestradiol. There was no clear relationship between the number of nigrosin-impermeable cells in suspensions prepared from different follicles and plating efficiency. It is concluded that the 3-h attachment of granulosa cells in culture is a useful measure of the number of viable cells in a suspension and is to be preferred to less direct techniques based on dye exclusion.
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D011863:Radioimmunoassay; D013739:Testosterone / Q000494:pharmacology
D013739:Testosterone; D004958:Estradiol