Gregoraszczuk E
Endocrinol Exp. 1983 Mar;17(1):59-68.
Porcine granulosa and luteum cells were isolated and cultured in vitro to compare their morphology and secretory activity with these of corpus luteum cells in vivo. Granulosa cells were isolated from preovulatory follicles and the luteal cells were collected from corpora lutea in the early or middle phase of the luteal cycle. The cells were grown as a monolayer for 8 days at 37 degrees C in the Parker's medium (M 199) supplemented with 10% of calf serum. The concentration of progestagens and estrogens in the medium was measured by the radioimmunological method [Abraham et al. 1971, Hotchkiss et al. 1971]. Both granulosa and luteal cells were cultured with the addition of following amounts of hormones: 100 ng LH, 600 ng hCG, 100 ng PRL and 150 ng estradiol per 1 ml of culture medium. At two days of culture the release of progesterone from granulosa cells significantly increased after the addition of 100 ng LH. Luteal cells isolated from the postovulatory corpus luteum released much higher amounts of progesterone than granulosa cells both in control cultures and in cultures supplemented with LH. A secretory stimulation of the luteal cells was observed only on the first day of culture irrespectively of the phase of the luteal cycle. In contrast, granulosa cells reacted by the elevation of hormonal release after the administration of hormones not only at the beginning of the culture, but also after several days. Prolactin did not stimulate the secretory activity of the granulosa cells and the same is valid for the cells isolated from the corpus luteum in the middle luteal phase. However, prolactin stimulated hormonal release of luteal cells isolated from the early corpus luteum.
分离猪的颗粒细胞和黄体细胞并进行体外培养,以将它们的形态和分泌活性与体内黄体细胞的形态和分泌活性进行比较。颗粒细胞从排卵前卵泡中分离出来,黄体细胞从黄体周期早期或中期的黄体中收集。细胞在补充有10%小牛血清的帕克培养基(M199)中于37℃作为单层培养8天。通过放射免疫法[亚伯拉罕等人,1971年;霍奇基斯等人,1971年]测量培养基中孕激素和雌激素的浓度。颗粒细胞和黄体细胞均在每1毫升培养基中添加以下量的激素进行培养:100纳克促黄体生成素(LH)、600纳克人绒毛膜促性腺激素(hCG)、100纳克催乳素(PRL)和150纳克雌二醇。在培养两天时,添加100纳克LH后,颗粒细胞中孕酮的释放显著增加。在对照培养物和补充有LH的培养物中,从排卵后黄体分离的黄体细胞释放的孕酮量均比颗粒细胞高得多。无论黄体周期处于哪个阶段,仅在培养的第一天观察到黄体细胞的分泌刺激。相比之下,颗粒细胞不仅在培养开始时,而且在数天后给予激素后,都会通过激素释放的升高做出反应。催乳素不会刺激颗粒细胞的分泌活性,对于从中期黄体分离的细胞也是如此。然而,催乳素会刺激从早期黄体分离的黄体细胞的激素释放。