梅花鹿卵巢颗粒细胞的体外培养与鉴定  

Culture and identification of granulosa cells from sika deer ovary

作　　者：邢明杰[1] 陈秀敏[1] 李晓霞[1] 刁云飞[1] 杨镒峰[1] 曹新燕[1] 王世勇[1] 赵伟刚[1] 许保增[1] 魏海军[1] 

机构地区：[1]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,吉林长春130112

出　　处：《中国兽医学报》2017年第6期1134-1138,共5页Chinese Journal of Veterinary Science

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)

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。Ovarian granulosa cells play an important role in the process of oocyte and follicular de- velopment. The cells not only provide nutrition for oocyte,but also secrete progesterone and estra- diol, and regulate the proliferation and differentiation of the follicle with ovarian membrane cell. O- varian granulosa cells have the stem cell properties, so they can be used as alternative to nuclear transfer. Follicle-stimulating hormone receptor antibody was used to identify granulosa cells easi- ly. In this research,sika deer granulose cells was cutured in vitro ,and aim at exploring the culture conditions of the sika deer granulosa cells for a long period, characterizing their morphology and phenotype. The results showed that sika deer ovarian granulosa cells showed polygon or fusi- formed, extended pseudopodia,and growed radiately. The granulosa cell purity was 〉90 % by the FSHR antibody tests. The sika deer ovarian granulosa cells cultured in DMEM/F12 nutrient solu tion containing 15% fetal bovine serum and 1% penicillin-streptomyc were good growth activity and high purity,easily identifed by HE staining and FSHR immunofluorescence staining.

关 键 词：梅花鹿 卵巢颗粒细胞 FSHR 体外培养 

分 类 号：Q343.6[生物学—遗传学]