Extracellular matrix regulates ovine granulosa cell survival, proliferation and steroidogenesis:: relationships between cell shape and function
Extracellular matrix regulates ovine granulosa cell survival, proliferation and steroidogenesis:: relationships between cell shape and function
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DOI：
10.1677/joe.0.1690347
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发表时间：
2001-05-01
影响因子：
4
通讯作者：
Monniaux, D
中科院分区：
文献类型：
--
作者：
Huet, C;Pisselet, C;Monniaux, D
细胞外基质（ECM）。构成卵泡基底层，也存在于卵泡之间和卵泡液中，被认为在卵泡发育过程中调节颗粒细胞（GC）的功能。从小（直径1-3毫米）或大（直径4-7毫米）的卵泡中分离出的绵羊GCs在各种纯ECM成分（I型胶原蛋白、纤维连接蛋白、层粘连蛋白）、增强（RGD肽）或损害（聚2-羟乙基甲基丙烯酸酯（聚血红素））细胞粘附的合成底物或肝素存在的情况下培养。这些因素单独使用或与IGF-I和/或FSH联合使用的影响，在GC扩散、存活、增殖和类固醇生成方面进行了评估。当生长在I型胶原蛋白（CI）凝胶，多血凝素或肝素。大小卵泡的GCs均呈圆形，增殖率低。与未包被塑料基质相比，这些ECM或合成化合物增加了大卵泡GCs的雌二醇分泌，减少了孕酮分泌。相比之下，来自大卵泡和小卵泡的GCs在CI涂层、纤维连接蛋白、层粘连蛋白和RGD肽上广泛扩散。纤维连接蛋白和层粘连蛋白显著提高了两种来源的GCs的增殖率和存活率。此外，纤维连接蛋白、层粘连蛋白和RGD肽减少了大卵泡GCs的雌二醇分泌。出乎意料的是，CI涂层增加了大卵泡GCs的雌二醇分泌，减少了孕酮分泌，这表明I型胶原能够独立于GC形状维持雌二醇分泌。最后，当细胞生长在ECM成分上时，GC对IGF-I和FSH的反应性，在增殖和甾体生成方面，通常保持不变。RGD肽和肝素的存在。然而，当大卵泡GCs作为非粘附簇生长时（如在多血瘤上观察到的），基础和IGF-I和/或fsh刺激的孕酮分泌完全被消除。总的来说，本研究表明GC形状。体外生存、增殖和甾体形成可以由纯ECM成分以特定和协调的方式调节。这表明，在体内，纤维连接蛋白和层粘连蛋白可能通过增强GCs的存活和增殖来维持卵泡发育，而I型胶原可能参与维持大窦卵泡的雌二醇分泌。
The extracellular matrix (ECM). constituting the follicular basal lamina and present also between follicular sells and in the follicular fluid, is believed to regulate granulosa cell (GC) function during follicular development. Ovine GCs isolated from small (1-3 mm in diameter) or large (4-7 mm in diameter) antral follicles were cultured on various pure ECM components (type I collagen, fibronectin, laminin), synthetic substrata enhancing (RGD peptides) or impairing (poly 2-hydroxyethylmethacrylate (poly-hema)) cell adhesion, or in the presence of heparin. The effects of these factors, used alone or in combination with IGF-I and/or FSH, were evaluated in terms of GC spread, survival, proliferation and steroidogenesis.When grown on type I collagen (CI) gel, poly-hema or heparin. GCs from both large and small follicles exhibited a round shape and a low proliferation rate. Compared with non-coated plastic substratum as a control, these ECM or synthetic compounds enhanced estradiol secretion and reduced progesterone secretion by large-follicle GCs. In contrast, GCs from both large and small follicles spread extensively on CI coating, fibronectin, laminin and RGD peptides. Fibronectin and laminin dramatically increased the proliferation rate and enhanced survival of GCs from both origins. Moreover, fibronectin, laminin and RGD peptides reduced estradiol secretion by large-follicle GCs. Unexpectedly, CI coating increased estradiol secretion and reduced progesterone secretion by large-follicle GCs, suggesting that type I collagen was able to maintain estradiol secretion independently of GC shape. Finally, GC responsiveness to IGF-I and FSH, in terms or proliferation and steroidogenesis, was generally maintained when cells were grown on ECM components. RGD peptides and in the presence of heparin. However, when large-follicle GCs were grown as non-adherent clusters (as observed on poly-hema) basal and IGF-I- and/or FSH-stimulated progesterone secretions were totally abolished.Overall, this study shows that GC shape. survival, proliferation and steroidogenesis can be modulated in vitro by pure ECM components in a specific and coordinated manner. It is suggested that, in vivo, fibronectin and laminin would sustain follicular development by enhancing the survival and proliferation of GCs, whereas type I collagen might participate in the maintenance of estradiol secretion in large antral follicles.