Development of a long-term bovine granulosa cell culture system: Induction and maintenance of estradiol production, response to follicle-stimulating hormone, and morphological characteristics
Development of a long-term bovine granulosa cell culture system: Induction and maintenance of estradiol production, response to follicle-stimulating hormone, and morphological characteristics
复制标题
DOI：
10.1095/biolreprod56.3.608
复制
发表时间：
1997-03-01
影响因子：
3.6
通讯作者：
Webb, R
中科院分区：
文献类型：
--
作者：
Gutierrez, CG;Campbell, BK;Webb, R
本研究的目的是建立一个无血清的牛颗粒细胞培养系统，在该系统中可以诱导和维持FSH反应性雌二醇的产生，并使用该系统来评估FSH、胰岛素和IGF-I对不同卵泡大小类别（直径< 4-、4-8-和> 8-mm）的牛颗粒细胞的类固醇生成和增殖的影响。在FSH存在下，来自小卵泡的颗粒细胞在体外分化，雌二醇分泌随时间增加（p < 0.01），因此在培养期结束时，其与来自大卵泡的细胞相似。中、大卵泡颗粒细胞在培养过程中均能分泌雌二醇。与无血清条件下培养的细胞相比，在血浆包被的培养威尔斯孔中培养的细胞具有增加的增殖反应，但具有较低的雌二醇产生（p < 0.01）。胰岛素促进三种卵泡大小的颗粒细胞增殖和雌二醇的产生（p < 0.01）。生理浓度的FSH以剂量反应方式诱导颗粒细胞增殖和雌二醇分泌（p < 0.01）。在培养体系中加入IGF-I可促进增殖和雌二醇的产生（p < 0.01），即使在缺乏促性腺激素支持的情况下也是如此，证明了这种生长因子的促性腺特性。该系统将允许控制牛颗粒细胞的分化和增殖的关键因素的详细研究。
The objectives of this study were to develop a serum-free bovine granulosa cell culture system in which FSH-responsive estradiol production could be induced and maintained, and to use this system to evaluate the effects of FSH, insulin, and IGF-I on steroidogenesis and proliferation of bovine granulosa cells from different follicle size categories (< 4-, 4-8-, and > 8-mm diameter). In the presence of FSH, granulosa cells from small follicles differentiated in vitro, and estradiol secretion increased with time (p < 0.01) so that by the end of the culture period it was similar to that of cells from large follicles. Granulosa cells from medium and large follicles secreted estradiol throughout the culture period. Cells cultured in plasma-coated culture wells had an increased proliferative response but had lower estradiol production compared to cells cultured under serum-free conditions (p < 0.01). Insulin promoted proliferation and estradiol production by granulosa cells from the three follicle-size categories (p < 0.01). Physiological concentrations of FSH induced proliferation and estradiol secretion (p < 0.01) by granulosa cells in a dose-responsive manner. The inclusion of IGF-I in the culture system enhanced proliferation and estradiol production (p < 0.01), even in the absence of gonadotropic support, demonstrating the gonadotropic characteristics of this growth factor.These results demonstrate the development of a relevant physiological culture system for bovine granulosa cells. This system will permit the detailed study of the key factors controlling the differentiation and proliferation of bovine granulosa cells.