中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (1): 78-84.doi: 10.3969/j.issn.2095-4344.2015.01.014

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

干细胞相关因子在大鼠卵巢颗粒细胞中的表达

刘慧萍，林政桦，张国民，喻  嵘，刘文娥，李  玲，谷旭宇   

  1. 湖南中医药大学中医方证研究转化医学实验室，湖南省长沙市  410208
  • 修回日期:2014-12-07 出版日期:2015-01-01 发布日期:2015-01-01
  • 通讯作者: 张国民，博士，副教授，硕士生导师，湖南中医药大学中医方证研究转化医学实验室，湖南省长沙市 410208
  • 作者简介:刘慧萍，女，博士，副教授，主要从事干细胞研究。
  • 基金资助:

    国家自然科学基金资助项目(81303123)；中国博士后基金项目(2013T60770，2012M511382)；高等学校博士学科点专项科研基金项目(20114323120007)；湖南省重点实验室开放基金(2014XNFZ06)

Granulosa cells with stem cell properties in the rat ovary

Liu Hui-ping, Lin Zheng-hua, Zhang Guo-min, Yu Rong, Liu Wen-e, Li Ling, Gu Xu-yu   

  1. Prescription and Syndrome Research and Translational Medicine Laboratory of Hunan University of Traditional Chinese Medicine, Changsha 410208, Hunan Province, China
  • Revised:2014-12-07 Online:2015-01-01 Published:2015-01-01
  • Contact: Zhang Guo-min, M.D., Associate professor, Master’s supervisor, Prescription and Syndrome Research and Translational Medicine Laboratory of Hunan University of Traditional Chinese Medicine, Changsha 410208, Hunan Province, China
  • About author:Liu Hui-ping, M.D., Associate professor, Prescription and Syndrome Research and Translational Medicine Laboratory of Hunan University of Traditional Chinese Medicine, Changsha 410208, Hunan Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81303123; China Postdoctoral Science Foundation, No. 2013T60770, 2012M511382; Specialized Research Fund for the Doctoral Program of Higher Education, No. 20114323120007; the Hunan Provincial Key Laboratory Foundation, No. 2014XNFZ06

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871

被引次数

1

摘要：

背景：现有研究表明，人与大鼠卵巢优势卵泡中的颗粒细胞具有表达干细胞特性的现象。
目的：探讨干细胞相关因子在大鼠卵巢颗粒细胞中的表达。
方法：将大鼠卵巢组织制作石蜡切片后，使用免疫组化法检测CD34，CD133，ABCG2/Bcrp1，Pou5f1/Oct-4的表达。卵泡穿刺法获取颗粒细胞，并使用免疫组化法检测FSHR受体的表达以鉴定颗粒细胞的纯度。免疫组化法检测颗粒细胞CD44，C-Kit的表达情况，RT-PCR检测卵巢组织与颗粒细胞ABCG2/Bcrp1、Pou5f1/Oct-4、Nanog基因的表达情况。
结果与结论：免疫组化法石蜡切片显示部分卵巢颗粒细胞CD34，CD133，ABCG2/Bcrp1，Pou5f1/Oct-4阳性表达，而Pou5f1/Oct-4蛋白的表达在卵泡发育的周期中逐步增多，并在黄体化的时候显著增强，形成白体后则消失，因而具有周期性表达的特点。卵泡穿刺法提取的原代颗粒细胞FSHR受体鉴定阳性率在95%以上。体外培养的颗粒细胞以长梭形或菱形为主，部分细胞有聚集生长现象，且CD44，C-Kit阳性表达。RT-PCR检测结果显示，卵巢组织与体外培养的颗粒细胞均不表达Nanog，卵巢低表达Pou5f1/Oct-4，强烈表达ABCG2/Bcrp1，颗粒细胞微弱表达Pou5f1/Oct-4，而ABCG2/Bcrp1表达较强。以上结果显示大鼠卵巢中的部分颗粒细胞具有干细胞的特性。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


全文链接：

关键词: 干细胞, 培养, 卵巢颗粒细胞, 干细胞样细胞, 干细胞特性, 集落现象, 阳性标志物, 体外培养, 国家自然科学基金

Abstract:

BACKGROUND: Human and rat ovarian granulosa cells in dominant follicles have the phenomenon of expressing stem cell characteristics.
OBJECTIVE: To investigate the expression of stem cell-related factors in rat ovarian granulosa cells.
METHODS: After the paraffin sections of rat ovarian tissue, immunohistochemical method was used to detect CD34, CD133, ABCG2/Bcrp1, Pou5f1/Oct-4 expressions. Granulosa cells cultured in vitro were harvested by follicular puncture method, and then the immunohistochemical method was used to detect the expression of FSHR receptor in order to identify the purity of granule cells. In the cultured granulosa cells, CD44 and C-Kit expressions were detected immunohistochemically, RT-PCR was used to detect ABCG2/Bcrp1, Pou5f1/Oct-4, Nanog gene expressions in ovarian tissue and granulosa cells.
RESULTS AND CONCLUSION: Immunohistochemistry detection on paraffin sections showed that a part of ovarian granulosa cells expressed CD34, CD133, ABCG2/Bcrp1 and Pou5f1/Oct-4, and the expression of Pou5f1/Oct-4 protein gradually increased in the development of ovarian follicles, significantly enhanced during the luteal phase, and then disappeared after the formation of corpus albicans, displaying a periodic expression characteristics. FSHR receptor positive identification rate of primary cells harvested by the folliclar puncture method was more than 95%. Granulosa cells cultured in vitro were mainly long spindle-shaped or diamond, and some cells presented with aggregation growth and expressed CD44 and C-Kit. RT-PCR test results showed that there were no Nanog in the ovarian tissue and cultured granulosa cells, low expression of Pou5f1/Oct-4 in the ovarian tissue, strong expression of ABCG2/Bcrp1 in the ovarian tissue, weak expression of Pou5f1/Oct-4 in the cultured granulosa cells, and strong expression of ABCG2/Bcrp1 in the cultured granulosa cells. These findings suggest that a part of granulosa cells in the rat ovarian have the characteristics of stem cells.


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


全文链接：

Key words: Stem Cells, Ovary, Granulosa Cells

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