Hillensjö T, Sjögren A, Strander B, Nilsson L, Wikland M, Hamberger L, Roos P
Acta Endocrinol (Copenh). 1985 Nov;110(3):401-7. doi: 10.1530/acta.0.1100401.
Granulosa cells were obtained from human preovulatory follicles in 31 women undergoing in vitro fertilization and embryo transfer due to tubal infertility. Follicular maturation was stimulated and synchronized by treatment with Clomiphene or human menopausal gonadotrophin (hMG), or both, plus human chorionic gonadotrophin (hCG). Follicles were aspirated by ultrasound guided puncture approximately 34-36 h after the hCG injection. The granulosa cells were washed and suspended in modified medium 199 containing 10% foetal bovine serum and cultured as monolayers for 6-8 days in the absence and presence of hormones and reactants. Progesterone formation was analyzed by RIA. In general, the cells underwent morphological luteinization and secreted high amount of progesterone. Under basal conditions the secretion of progesterone was highest during the first 2 days in culture and then gradually declined. Progesterone secretion was stimulated by human LH, hCG and the adenylate cyclase stimulator forskolin, with a maximal effect between days 2-6. The beta-adrenergic agonist isoproterenol in preliminary experiments potentiated the stimulatory effect of hCG but had no own stimulatory effect. No clear differences in progesterone secretion or responsiveness to in vitro stimulation relating to the various in vivo stimulation protocols were found.
从31名因输卵管性不孕而接受体外受精和胚胎移植的女性的人排卵前卵泡中获取颗粒细胞。通过使用克罗米芬或人绝经期促性腺激素（hMG）或两者加人绒毛膜促性腺激素（hCG）进行治疗来刺激和同步卵泡成熟。在注射hCG后约34 - 36小时，通过超声引导穿刺抽吸卵泡。将颗粒细胞洗涤并悬浮于含有10%胎牛血清的改良培养基199中，在无激素和反应物以及有激素和反应物的情况下单层培养6 - 8天。通过放射免疫分析法分析孕酮的生成。一般来说，细胞经历形态学上的黄体化并分泌大量孕酮。在基础条件下，孕酮分泌在培养的头2天最高，然后逐渐下降。人促黄体生成素（LH）、hCG和腺苷酸环化酶刺激剂福斯高林刺激孕酮分泌，在第2 - 6天之间有最大效应。在初步实验中，β - 肾上腺素能激动剂异丙肾上腺素增强了hCG的刺激作用，但自身没有刺激作用。未发现与各种体内刺激方案相关的孕酮分泌或对体外刺激反应性方面的明显差异。
