SURVIVAL AND GONADOTROPHIN RESPONSIVENESS OF LUTEAL CELLS IN VITRO
ABSTRACT Simplified methods for incubating luteal tissues have been developed. Progesterone biosynthesis in washed, minced bovine luteal tissue is stimulated by added bovine LH. The response is linear in the range 2–200 ng added LH per ml of medium. The use of covariance analysis to correct for differences in time elapsing between mincing the tissue and the beginning of incubation reduces the standard error of the mean and results in marked improvement in the indices of precision. Progesterone biosynthesis in washed, minced bovine luteal tissues appears specific for LH; no other pituitary hormones give a response, and the response is completely negated by adding anti-bovine LH serum to the incubation medium. Prior treatment of cattle from which the incubated corpora lutea are obtained with various levels of human chorionic gonadotrophin, results in greatly increased luteal tissue weights. However, this tissue has a markedly reduced sensitivity to LH added in vitro. No conclusive evidence was found for a feedback inhibition of progesterone on its own synthesis in this system. The incubation system developed is sensitive enough to serve as a bioassay for LH in biological tissues and fluids. Preliminary data suggest that a simplified protein binding assay can be successfully used to measure the progestins synthesized by minced, washed bovine luteal tissue in response to added bovine LH.