Characteristics of bovine luteal cells in culture: morphology, proliferation and progesterone secretion in different media and effects of LH, dibutyryl cyclic AMP, antioxidants and insulin.

Abstract

The morphological and functional characteristics of cultured bovine luteal cells were examined. Dispersed luteal cells were cultured in either Ham's F12 medium or Dulbecco's Modified Eagle's Medium (DMEM), plus 10% adult bovine serum. After 18 h of culture, fibronectin was observed to be associated with some cells showing histochemical 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity. In DMEM, cells adopted a fibroblast-like appearance with a doubling time of 34 h while in Ham's F12, cells retained an epithelial-like morphology with a doubling time of 91 h. Dibutyryl cyclic AMP (dbcAMP) but not LH slowed proliferation and altered cell morphology in DMEM. Addition of insulin and dbcAMP was required to maintain 3 beta-HSD activity in cells cultured for more than 3 days. Progesterone secretion was significantly greater in Ham's F12 than in DMEM although there was a rapid decline in basal secretion during culture in either medium. Both LH and dbcAMP stimulated progesterone secretion although the effects of dbcAMP were significantly greater than those of LH after 18 h of culture. Addition of insulin significantly enhanced basal, LH-stimulated and dbcAMP-stimulated progesterone secretion after 3 days of culture whereas several antioxidants were without effect. These studies indicate that the morphological and functional characteristics of cultured luteal cells may be best maintained in Ham's F12 plus dbcAMP and insulin.