摘要
[目的]建立一种高效的水牛子宫内膜腺上皮细胞和基质细胞分离培养方法,为胚胎着床和子宫内膜相关疾病的分子生物学机制研究奠定基础。[方法]采用酶消化、刮取、系列过滤和差速贴壁相结合的方法分离水牛子宫内膜腺上皮细胞和基质细胞,用免疫细胞化学法和台盼兰染色法检测分离细胞的纯度和活率。[结果]成功分离培养出子宫内膜腺上皮细胞和基质细胞;免疫荧光染色和细胞计数表明上皮细胞和基质细胞的纯度均达90%以上;台盼兰染色结果显示,上皮细胞的活率为91%,基质细胞的活率为78%。[结论]采用酶消化、刮取、过滤和差速贴壁相结合的方法可分离出高纯度的水牛子宫内膜腺上皮细胞和基质细胞。
[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells （BESC） and endometrial glandular epithelial ceils（ BEGEC ） of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [ Method ] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated ceils. [ Result]The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purification was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [ Conclusion ]The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purification.
出处
《安徽农业科学》
CAS
北大核心
2009年第18期8512-8514,8517,共4页
Journal of Anhui Agricultural Sciences
基金
广西大学研究生创新项目(2008105930905D001)
广西科技厅科技攻关项目(桂科攻0815008-2-4)
