Bovine luteal cells in tissue culture

Abstract

Luteal cells obtained from bovine corpus luteum were grown in tissue culture in a medium consisting of Dulbecco's modified Eagle supplemented with 10% horse serum. The cells grew under these conditions for at least 30 days. When the cells were seeded at low density they grew in an epithelioid pattern. After high density seeding they took a fibroblast-like appearance. The cells showed a lacy cytoplasm with vacuoles of various sizes, which contained material identified as lipid by staining with Sudan Black B and Oil Red O and they do react strongly for 3β hydroxysteroid dehydrogenase and glucose-6-phosphate dehydrogenase. They secreted progesterone and 20α Hydroxy pregn-4-en-3-one, and were able to metabolize pregnenolone into progesterone and 20α Hydroxy pregn-4-en-3-one at a high rate. These two steroids were produced in a ratio similar to that observed in vivo. When the luteal cells were cultured in the presence of luteinizing hormone (LH), the daily secretion of progestin decreased slowly over a period of 14 days. In the absence of LH, the rate of progestin secretion diminished drastically during the first week in culture, and the subsequent addition of LH caused only a slight stimulation of progestin secretion as compared to the rate of secretion of cells cultured from the start in the presence of LH.