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首页> 外文期刊>Comparative biochemistry and physiology, Part C. Pharmacology, toxicology and endocrinology: An international journal >Basal lamina of avian ovarian follicle: influence on morphology of granulosa cells in-vitro
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Basal lamina of avian ovarian follicle: influence on morphology of granulosa cells in-vitro

机译：禽卵泡基底层：对体外颗粒细胞形态的影响

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Experiments were conducted to determine the influence of basal lamina on the morphology of ovarian granulosa. cells in vitro. Pure and intact basal lamina was isolated from the large preovulatory follicles of the chicken ovary and designated basal lamina of avian ovarian follicle (BLAOF). Examination of the isolated basal lamina with electron microscope revealed an ultrastructure that is similar to that of basal lamina in the intact ovarian follicle. Pieces of the intact basal lamina were attached to the bottom of 32 mm culture dishes (BLAOF-coated dishes) in which differentiated granulosa cells isolated from the largest preovulatory follicle or undifferentiated granulosa cells isolated from immature small yellow chicken ovarian follicles were cultured; uncoated dishes served as controls. Granulosa cells incubated on intact basal lamina assumed spherical shape, whereas granulosa cells incubated directly on plastic in control dishes became highly flattened. Interestingly, granulosa cells that attached to plastic close to BLAOF (in BLAOF-containing dishes) became rounded. The storage of BLAOF-coated culture dishes at 4 degrees C for 2 years had no apparent effect on its ability of the matrix material to induce changes in granulosa cell shape. Some components of the basal lamina could be solubilized with guanidine-HCl alone (fraction 1; 90-95% of total protein in BLAOF) with the remaining components solubilized with P-mercaptoethanol containing guanidine-HCl (fraction 2; 5-10% of total protein in BLAOF). Differentiated and undifferentiated chicken granulosa cells became rounded when incubated in fraction 1-pre-coated wells: whereas those incubated directly on plastic in control wells were flattened. Similarly, when fraction 1 of solubilized basal lamina was added as liquid to incubation mixture, it caused both differentiated and undifferentiated granulosa cells to assume spherical shapes. The storage of fraction 1-coated culture dishes at 4 degrees C for 12 or more months had no apparent effect on its ability to influence granulosa cell shape. Fraction 1-induced changes in granulosa cell shape were similar to those observed for complete and intact basal lamina (BLAOF). These findings demonstrate that intact homologous basal lamina (BLAOF) or its solubilized (fluidized) form can induce normal (in vivo) morphology in granulosa cells. It is suggested that BLAOF or its solubilized form can be used to culture cells in experiments designed to examine the influence of the natural basal lamina microenvironment on cellular behavior and function. (C) 2000 Elsevier Science Inc. All rights reserved. [References: 22]
机译：进行实验以确定基底层对卵巢颗粒形态的影响。体外细胞。从鸡卵巢的大排卵前卵泡中分离出纯净而完整的基底层，并将其命名为禽卵泡的基底层（BLAOF）。用电子显微镜检查孤立的基底层，发现其超微结构类似于完整卵巢卵泡中基底层的超微结构。将完整的基底叶基片附着在32毫米培养皿（涂有BLAOF的培养皿）的底部，在其中培养从最大排卵前卵泡分离的分化颗粒细胞或从未成熟的黄色小鸡卵巢卵泡分离的未分化颗粒细胞。将未涂层的菜肴作为对照。在完整的基底层上孵育的颗粒细胞呈球形，而在对照培养皿中直接在塑料上孵育的颗粒细胞高度扁平化。有趣的是，附着在靠近BLAOF的塑料上的颗粒细胞（在含BLAOF的培养皿中）变得圆形。 BLAOF包被的培养皿在4摄氏度下保存2年，对基质材料诱导颗粒细胞形状变化的能力没有明显影响。基底层的某些成分可以单独用盐酸胍盐溶解（级分1; BLAOF中总蛋白质的90-95％），其余成分可以用含胍盐酸盐的P-巯基乙醇溶解（级分2; 5-10％ BLAOF中的总蛋白质）。当在组分1预包被的孔中孵育时，分化的和未分化的鸡颗粒细胞变得圆形：而在对照孔中直接在塑料上孵育的细胞则变平。类似地，当将溶解的基底层的级分1作为液体添加到孵育混合物中时，它会导致分化和未分化的颗粒细胞呈球形。将涂有级分1的培养皿在4摄氏度下存放12个月或更长时间，对其影响颗粒细胞形状的能力没有明显影响。分数1诱导的颗粒细胞形状变化与完整和完整的基底层（BLAOF）观察到的相似。这些发现表明，完整的同源基底膜（BLAOF）或其增溶（流化）形式可在颗粒细胞中诱导正常（体内）形态。建议在实验中使用BLAOF或其可溶形式培养细胞，以检查天然基底薄层微环境对细胞行为和功能的影响。 （C）2000 Elsevier Science Inc.保留所有权利。 [参考：22]

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