LH release from dispersed bovine pituitary cells in culture: in vitro effects of estradiol and procedural variables.
Procedures for cell dissociation and in vitro culture were validated to investigate secretion of luteinizing hormone (LH) from bovine anterior lobe (AL) pituitary cells. The concentration of trypsin used for dissociation affected cell yield, cell loss during preincubation, LH secretion, and response to gonadotropin-releasing hormone (GnRH). Optimum results were obtained with trypsin concentrations of 8-16 micrograms/mg fresh tissue. Duration of preincubation and of experimental culture markedly affected LH secretion and response to GnRH. Immediately after dissociation, cells contained relatively low quantities of LH, but they were able to release a substantial proportion of this LH. Basal release of LH and GnRH-induced release of LH were highly correlated with total quantities of LH, and all three parameters increased with time of preincubation until 24 hr. Experimental treatments of 2 hr duration were optimal for investigating GnRH stimulation of LH release, whereas longer treatments may be required to investigate effects of agents that inhibit the release of LH. Preincubation of dissociated AL cells with physiological concentrations of estradiol increased all three LH parameters. Progesterone had no effect either alone or in combination with estradiol. In conclusion, the procedures described for cell dissociation and culture of suspended cells provide a useful tool for studying release of LH from the bovine AL cell.
D000818:Animals; D002417:Cattle / Q000502:physiology*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004958:Estradiol / Q000494:pharmacology*; D007986:Luteinizing Hormone / Q000378:metabolism*; D008297:Male; D010903:Pituitary Gland, Anterior / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011374:Progesterone / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
D011374:Progesterone; D004958:Estradiol; D007986:Luteinizing Hormone; D014357:Trypsin