Sites C K, Kessel B, LaBarbera A R
Department of Obstetrics and Gynecology, University of Cincinnati College of Medicine, Ohio 45267-0526, USA.
Proc Soc Exp Biol Med. 1996 May;212(1):78-83. doi: 10.3181/00379727-212-43994.
We sought to determine the influence of different constituents of the extracellular matrix on porcine granulosa cell function by assessing cellular attachment, cellular morphology, follicle-stimulating hormone (FSH) receptors, and progesterone production. Cells from immature porcine ovarian follicles were cultured for up to 6 days in serum-free medium containing porcine FSH (pFSH, 10 ng/ml) in culture dishes either uncoated or coated with one of the following adhesion proteins: gelatin (1 mg/cm2), fibronectin (1 microgram/cm2), laminin (1 microgram/cm2), type I collagen (10 micrograms/cm2), or type IV collagen (7.8 micrograms/cm2). Fibronectin, laminin, type I collagen, and type IV collagen increased cellular attachment significantly (P < 0.05). All adhesion proteins except gelatin influenced cellular morphology. Cells cultured on laminin or type IV collagen formed dense clusters of rounded cells. Cells cultured in dishes coated with each adhesion protein except gelatin had higher 125I-pFSH binding per cell than cells cultured in uncoated dishes, with increases of 7- to 12-fold over control (P < 0.05). All adhesion proteins increased progesterone production, ranging from 10- to 50-fold over control (P < 0.05). In summary, not only did adhesion proteins increase attachment to the dishes but they also increased FSH receptors and differentiated function (progesterone production) of granulosa cells from immature porcine ovarian follicles.
我们试图通过评估细胞黏附、细胞形态、促卵泡激素(FSH)受体和孕酮生成,来确定细胞外基质的不同成分对猪颗粒细胞功能的影响。将来自未成熟猪卵泡的细胞在含有猪FSH(pFSH,10 ng/ml)的无血清培养基中,于未包被或包被有下列黏附蛋白之一的培养皿中培养长达6天:明胶(1 mg/cm²)、纤连蛋白(1 μg/cm²)、层粘连蛋白(1 μg/cm²)、I型胶原(10 μg/cm²)或IV型胶原(7.8 μg/cm²)。纤连蛋白、层粘连蛋白、I型胶原和IV型胶原显著增加了细胞黏附(P < 0.05)。除明胶外,所有黏附蛋白均影响细胞形态。在层粘连蛋白或IV型胶原上培养的细胞形成了密集的圆形细胞簇。在除明胶外的每种黏附蛋白包被的培养皿中培养的细胞,其每细胞的¹²⁵I-pFSH结合量高于未包被培养皿中的细胞,比对照增加了7至12倍(P < 0.05)。所有黏附蛋白均增加了孕酮生成,比对照增加了10至50倍(P < 0.05)。总之,黏附蛋白不仅增加了细胞对培养皿的黏附,还增加了未成熟猪卵泡颗粒细胞的FSH受体和分化功能(孕酮生成)。