A Technique for the Isolation of Bovine Luteal Cells and Its Application to Metabolic Studies of Luteal Cellsin Vitro

This is the authors' abstract. We don't add key points for this paper.

Homogeneous populations of luteal cells were obtained from slices of bovine corpus luteum by treatment with a crude preparation of collagenase and hyaluronidase followed by trypsin and DNAse. Cell recovery was about 20% as determined by protein and DNA assays. Viability of recovered cells was greater than 90% as judged by the trypan blue exclusion test. No gross change in cellular structure took place during the isolation procedure and the cell membrane remained intact. The free cells were able to incorporate 3H leucine into proteins at a constant rate for at least 18 hr. The isolated luteal cells were sensitive to luteinizing hormone (LH). In the presence of LH, 3.3 × 10-13M (10 pg/ml), the secretion of progestins was measurably enhanced. The secretion was linear as a function of time (3 hr in Krebs-Ringer bicarbonate solution) and maximal stimulation was obtained at 3.3 × 10-9M (100 ng/ml). This preparation can thus be used to study the long term effects of LH on the metabolism of luteal cells in vitro. (Endocrinology90: 1427, 1972)