Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan.
Endocrinology. 2012 Jun;153(6):2851-60. doi: 10.1210/en.2011-1810. Epub 2012 Mar 30.
The ovary is a complex endocrine organ responsible for steroidogenesis and folliculogenesis. Follicles consist of oocytes and two primary steroidogenic cell types, the granulosa cells, and the theca cells. Immortalized human granulosa cells are essential for researching the mechanism of steroidogenesis and folliculogenesis. We obtained granulosa cells from a 35-yr-old female and immortalized them by lentivirus-mediated transfer of several genes so as to establish a human nonluteinized granulosa cell line (HGrC1). We subsequently characterized HGrC1 and investigated its steroidogenic performance. HGrC1 expressed enzymes related to steroidogenesis, such as steroidogenic acute regulatory protein, CYP11A, aromatase, and gonadotropin receptors. Stimulation with FSH increased the mRNA levels of aromatase, which consequently induced the aromatization of androstenedione to estradiol. Activin A increased the mRNA levels of the FSH receptor, which were synergistically up-regulated with FSH stimulation. HGrC1 also expressed a series of ligands and receptors belonging to the TGF-β superfamily. A Western blot analysis showed that bone morphogenetic protein (BMP)-4, BMP-6, and BMP-7 phosphorylated small mother against decapentaplegic (Smad)1/5/8, whereas growth differentiation factor-9 phosphorylated Smad2/3. BMP-15 and anti-Müllerian hormone phosphorylated Smad1/5/8 while also weakly phosphorylating Smad2/3. These results indicate that HGrC1 may possess the characteristics of granulosa cells belonging to follicles in the early stage. HGrC1 might also be capable of displaying the growth transition from a gonadotropin-independent status to gonadotropin-dependent one.
卵巢是一个复杂的内分泌器官，负责甾体生成和卵泡发生。卵泡由卵母细胞和两种主要的甾体生成细胞类型，即颗粒细胞和膜细胞组成。永生化的人颗粒细胞对于研究甾体生成和卵泡发生的机制至关重要。我们从一位 35 岁的女性获得颗粒细胞，并通过慢病毒介导的几种基因转移使其永生化，从而建立了一种人未黄体化颗粒细胞系（HGrC1）。随后，我们对 HGrC1 进行了表征，并研究了其甾体生成性能。HGrC1 表达与甾体生成相关的酶，如甾体生成急性调节蛋白、CYP11A、芳香酶和促性腺激素受体。FSH 的刺激增加了芳香酶的 mRNA 水平，进而诱导雄烯二酮向雌二醇的芳香化。激活素 A 增加了 FSH 受体的 mRNA 水平，与 FSH 刺激协同上调。HGrC1 还表达一系列属于 TGF-β超家族的配体和受体。Western blot 分析表明，骨形态发生蛋白 4、6 和 7 磷酸化 small mother against decapentaplegic（Smad）1/5/8，而生长分化因子 9 磷酸化 Smad2/3。BMP-15 和抗苗勒管激素磷酸化 Smad1/5/8，同时也弱磷酸化 Smad2/3。这些结果表明，HGrC1 可能具有早期卵泡颗粒细胞的特征。HGrC1 也可能能够表现出从促性腺激素非依赖性状态向促性腺激素依赖性状态的生长转变。
