Vanderstichele H, Delaey B, de Winter J, de Jong F, Rombauts L, Verhoeven G, Dello C, van de Voorde A, Briers T
Innogenetics N.V., Ghent, Belgium.
Biol Reprod. 1994 May;50(5):1190-202. doi: 10.1095/biolreprod50.5.1190.
The expression and function of gonadotropin receptors, and the secretion of steroids, transferrin, and cytokines were investigated in three immortalized (single transfection with v-myc) mouse granulosa cell lines (GRM01, GRM01L, and GRM02). A dose-dependent increase in progesterone production was obtained in GRM01 and GRM02 cells after addition of LH, FSH, modulators of the adenylate cyclase enzyme system, and cAMP analogues. The LH-induced release of progesterone was already detectable in GRM02 cells after 8 h and was related to incubation time and cell number. Both epidermal growth factor (EGF) and transforming growth factor alpha (TGF alpha) induced the secretion of progesterone in GRM02 cells, while no effect was obtained with TGF beta. LH receptor concentration was highest in the GRM02 cell line. FSH receptor mRNA was visualized in GRM01 and GRM02 cells. Aromatase activity in GRM02 cells was induced by androgens and inhibited by aromatase inhibitors. Whereas all cell lines were able to secrete transferrin, only in GRM01 cells was transferrin secretion increased significantly by LH. FSH did not affect transferrin secretion in the three cell lines, in contrast to forskolin or 8-bromo-cAMP. The immortalized mouse granulosa cell lines were able to express and release several growth factors. The expression and secretion of activin, inhibin, TGF beta, EGF, TGF alpha, insulin-like growth factor II, fibroblast growth factor (acidic and basic), platelet-derived growth factor, and interleukin-6 suggest an autocrine or paracrine role for these factors in follicular differentiation and function. In conclusion, these cells, derived from mural granulosa cells and immortalized in a preovulatory state, can be used to study granulosa cell physiology or to study the role of granulosa cells and their derivatives in the process of follicular maturation, fertilization, and early embryonic development.
在三种永生化（用v-myc单转染）小鼠颗粒细胞系（GRM01、GRM01L和GRM02）中，研究了促性腺激素受体的表达和功能，以及类固醇、转铁蛋白和细胞因子的分泌。在添加促黄体生成素（LH）、促卵泡生成素（FSH）、腺苷酸环化酶酶系统调节剂和环磷酸腺苷（cAMP）类似物后，GRM01和GRM02细胞中的孕酮产量呈剂量依赖性增加。在GRM02细胞中，8小时后即可检测到LH诱导的孕酮释放，且与孵育时间和细胞数量有关。表皮生长因子（EGF）和转化生长因子α（TGFα）均诱导GRM02细胞分泌孕酮，而转化生长因子β（TGFβ）则无此作用。GRM02细胞系中的LH受体浓度最高。在GRM01和GRM02细胞中可观察到FSH受体mRNA。GRM02细胞中的芳香化酶活性受雄激素诱导，并被芳香化酶抑制剂抑制。虽然所有细胞系都能分泌转铁蛋白，但只有GRM01细胞中的转铁蛋白分泌因LH而显著增加。与福斯可林或8-溴-cAMP不同，FSH对这三种细胞系中的转铁蛋白分泌没有影响。永生化小鼠颗粒细胞系能够表达和释放多种生长因子。激活素、抑制素、TGFβ、EGF、TGFα、胰岛素样生长因子II、成纤维细胞生长因子（酸性和碱性）、血小板衍生生长因子和白细胞介素-6的表达和分泌表明，这些因子在卵泡分化和功能中具有自分泌或旁分泌作用。总之，这些源自壁层颗粒细胞并在排卵前状态下永生化的细胞，可用于研究颗粒细胞生理学，或研究颗粒细胞及其衍生物在卵泡成熟、受精和早期胚胎发育过程中的作用。
