Rodgers R J, O'Shea J D, Findlay J K
J Reprod Fertil. 1983 Sep;69(1):113-24. doi: 10.1530/jrf.0.0690113.
Corpora lutea from cyclic ewes were dissociated by collagenase digestion and trypsin/EGTA treatment. Enriched fractions of endothelial cells, small luteal cells and large luteal cells were prepared on a stepped gradient of Ficoll 400. Progesterone was measured by radioimmunoassay and the results corrected so that progesterone production by each cell type could be determined. Endothelial cells did not produce significant amounts of progesterone, with or without LH stimulation, and endothelial cell contamination of small and large luteal cell fractions did not influence progesterone production by these fractions. Mean +/- s.e.m. basal progesterone production (n = 10) by large luteal cells was greater (P less than 0.001) on a per cell basis than that by small luteal cells (1.16 +/- 0.16 compared with 0.25 +/- 0.06 pg/h/cell). However LH, which stimulated a maximal 3-4-fold increase in progesterone production by small luteal cells (LH ED50 = 0.14 ng/ml), had no significant effect on production by large luteal cells, when contamination by small luteal cells was taken into account. The response of small luteal cells was specific to LH, other hormones having had no significant effect. Basal progesterone production by small luteal cells (0.12 +/- 0.03 fg/h/micron3) calculated per unit volume of cell was not significantly different from that of large luteal cells (0.17 +/- 0.02 fg/h/micron3). After LH stimulation, small luteal cells produced more progesterone than did large luteal cells (0.40 +/- 0.09 compared with 0.18 +/- 0.03 fg/h/micron3) (P less than 0.05). When the amounts of progesterone produced per cell were multiplied by the absolute numbers of large luteal (1 X 10(7] and small luteal (5 X 10(7] cells in the intact corpus luteum, basal progesterone production by large luteal cells (11.6 +/- 1.6 micrograms/h) was similar to that by small luteal cells (12.3 +/- 3.0 micrograms/h). However, under LH stimulation, progesterone production by the small luteal cell type (39.9 +/- 9.5 micrograms/h) was approximately 3 times greater than that by the large luteal cell type (12.3 +/- 1.6 micrograms/h) (P less than 0.05). We therefore conclude that small luteal cells may be the principal source of luteal progesterone production in the sheep.
通过胶原酶消化和胰蛋白酶/乙二醇双乙酸酯处理对处于发情周期的母羊的黄体进行解离。在菲可400的阶梯梯度上制备富含内皮细胞、小黄体细胞和大黄体细胞的组分。通过放射免疫测定法测量孕酮，并对结果进行校正，以便能够确定每种细胞类型的孕酮生成量。无论有无促黄体生成素（LH）刺激，内皮细胞都不会产生大量孕酮，并且小黄体细胞和大黄体细胞组分中的内皮细胞污染不会影响这些组分的孕酮生成。以每个细胞为基础，大黄体细胞的平均±标准误基础孕酮生成量（n = 10）比小黄体细胞的更高（P < 0.001）（分别为1.16 ± 0.16与0.25 ± 0.06 pg/h/细胞）。然而，考虑到小黄体细胞的污染情况，刺激小黄体细胞孕酮生成量最大增加3 - 4倍的LH（LH半数有效剂量 = 0.14 ng/ml）对大黄体细胞的生成量没有显著影响。小黄体细胞的反应对LH具有特异性，其他激素没有显著影响。按单位细胞体积计算，小黄体细胞的基础孕酮生成量（0.12 ± 0.03 fg/h/μm³）与大黄体细胞的基础孕酮生成量（0.17 ± 0.02 fg/h/μm³）没有显著差异。在LH刺激后，小黄体细胞产生的孕酮比大黄体细胞更多（分别为0.40 ± 0.09与0.18 ± 0.03 fg/h/μm³）（P < 0.05）。当将每个细胞产生的孕酮量乘以完整黄体中大黄体细胞（1×10⁷）和小黄体细胞（5×10⁷）的绝对数量时，大黄体细胞的基础孕酮生成量（11.6 ± 1.6 μg/h）与小黄体细胞的基础孕酮生成量（12.3 ± 3.0 μg/h）相似。然而，在LH刺激下，小黄体细胞类型的孕酮生成量（39.9 ± 9.5 μg/h）大约是大黄体细胞类型（12.3 ± 1.6 μg/h）的3倍（P < 0.05）。因此，我们得出结论，小黄体细胞可能是绵羊黄体中孕酮生成的主要来源。
