梅花鹿卵巢颗粒细胞的体外培养与鉴定

发布时间：2018-03-31 08:36

本文选题：梅花鹿　切入点：卵巢颗粒细胞　出处：《中国兽医学报》2017年06期

【摘要】：卵巢颗粒细胞在卵母细胞生长和卵泡发育过程中起着重要作用,为卵母细胞提供营养,分泌孕酮和雌二醇,与膜细胞一起调控卵泡的增殖和分化。卵巢颗粒细胞也具有干细胞特性,可以作为核移植的替代物,用卵泡刺激素受体(FSHR)抗体能很快地鉴定颗粒细胞。本研究通过对梅花鹿卵巢颗粒细胞进行体外培养,旨在探索梅花鹿颗粒细胞长期体外培养的条件,描述颗粒细胞的形态和表型特征,为进一步研究颗粒细胞的功能奠定基础。试验结果表明,梅花鹿卵巢颗粒细胞呈多角形或梭形,并伸出伪足,呈放射状生长,FSHR抗体鉴定结果为颗粒细胞纯度90%。用含15%胎牛血清和1%双抗的DMEM/F12培养液培养,可获得生长活性好、细胞纯度高的梅花鹿卵巢颗粒细胞,HE染色和FSHR免疫荧光染色可以简便快速地鉴定梅花鹿卵巢颗粒细胞。
[Abstract]:Ovarian granulosa cells play an important role in oocyte growth and follicle development, providing nutrients for oocytes and secreting progesterone and estradiol. Ovarian granulosa cells also have stem cell properties and can be used as a substitute for nuclear transplantation. Granulosa cells could be quickly identified by FSHR-antibody. In order to explore the conditions for long-term culture of sika deer granulosa cells in vitro, the granulosa cells of sika deer ovary were cultured in vitro. The morphological and phenotypic characteristics of granulosa cells were described in order to lay a foundation for further study on the function of granulosa cells. The results showed that granulosa cells of sika deer ovary were polygonal or fusiform and outstretched pseudopodia. The results of FSHR antibody identification showed that the purity of granulosa cells was 90%. The growth activity could be obtained by using DMEM/F12 medium containing 15% fetal bovine serum and 1% double antibody. High purity sika deer ovarian granulosa cells were identified by HE staining and FSHR immunofluorescence staining.
【作者单位】：中国农业科学院特产研究所特种经济动物分子生物学重点实验室;
【基金】：中国农业科学院基本科研业务费增量资助项目(2015ZL012);中国农业科学院科技创新工程资助项目(CAAS-ASTIP-2015-ISAPS)
【分类号】：S825

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