南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (07): 914-917.

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人绒毛外细胞滋养层细胞与蜕膜基质细胞共培养模型的建立

秦立赟； 陈士岭； 张曦倩； 王炎秋；   

  1. 南方医科大学南方医院妇产科生殖医学中心； 南方医科大学南方医院妇产科生殖医学中心 广东广州510515； 广东广州510515；
  • 出版日期:2006-07-20 发布日期:2006-07-20
  • 基金资助:
    国家自然科学基金(30470657);广东省自然科学基金(04020416)~~

Co-culture of human extravillous cytotrophoblasts and decidual stromal cells in vitro

QIN Li-yun, CHEN Shi-ling, ZHANG Xi-qian, WANG Yan-qiu Center for Reproductive Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China   

  1. 南方医科大学南方医院妇产科生殖医学中心； 南方医科大学南方医院妇产科生殖医学中心 广东广州510515； 广东广州510515；
  • Online:2006-07-20 Published:2006-07-20

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摘要： 目的模拟体内环境,建立可保持绒毛外细胞滋养层细胞(EVCT)与蜕膜基质细胞(DSC)生物学特性的共培养细胞模型,应用于研究滋养细胞的侵袭行为。方法利用胰酶消化和BSA、Percoll梯度沉降,收集纯化人早孕绒毛组织的EVCT和DSC,分别放入Transwell的上下小室,观察该模型下细胞形态变化及侵袭特性,免疫组化、免疫荧光染色检测细胞的纯度。结果免疫组化显示EVCT的细胞角蛋白7阳性表达的细胞数占95%以上,阳性表达转化生长因子β2的细胞数占95%以上,DSC泌乳素阳性表达的细胞数也超过95%,证实共培养系统中EVCT和DSC纯度均在95%以上,且生物学特性得以维持。上室中的EVCT保持了其侵袭能力,在Matrigel包被的滤膜上EVCT的侵袭指数为3.22±0.04。结论适当的培养方法可获得高纯度的EVCT和DSC,并使其维持特有的生物学活性,成功地建立了共培养系统模型,便于研究滋养细胞侵袭和胚胎着床的机制。 

Abstract: Objective To develop a convenient method for isolation and purification of human extravillous cytotrophoblasts (EVCTs) and decidual stromal cells (DSCs) and establish a co-culture system. Methods The DSCs were digested with trypsin and purified by Percoll gradient. The EVCTs were digested with trypsin and purified by BSA gradient. Immunohischemistry and immunofluorescent study are performed to characterize these isolated cells. The EVCTs and DSCs were placed in Matrigel-coated Transwell upper and lower chamber, respectively, to study the invasive ability of the EVCTs. Results Immunohischemistry revealed that the purity of EVCTs and DSC exceeded 95%. Cultured EVCTs retained their capacity to invade Matrigel-coated Transwell filters with the invasion index of 3.22±0.04. Conclusion This co-culture modelestablished by isolating highly purified EVCTs and DSCs in vitro can be useful for studying the trophoblast invasion mechanisms .

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