不同分离纯化法建立人早孕滋养细胞模型的效果比较

发布时间：2018-03-01 11:07

本文关键词： 早孕滋养细胞原代培养胎盘人外周血淋巴细胞分离液 percoll密度梯度差速细胞角蛋白-　出处：《广东医学》2015年05期 　论文类型：期刊论文

【摘要】：目的对4种建立滋养细胞的方法进行比较,以期寻求一种简便高效的人早孕滋养细胞的体外培养方法,为相关研究提供基础。方法早孕绒毛通过胰酶联合胶原酶消化分离后,分别采用直接接种、差速贴壁联合差速消化法、人外周血淋巴细胞分离液分离纯化法、完整绒毛消化联合简化的percoll密度梯度分离法4种方法纯化培养,倒置显微镜观察细胞形态,应用HE染色、免疫组化和免疫细胞化学法检测细胞纯度。结果直接接种、差速贴壁联合差速消化法、人外周血淋巴细胞分离液分离纯化法得到的细胞细胞角蛋白7(CK-7)表达阳性率不足60%;简化的percoll密度梯度分离法得到的细胞CK-7表达阳性率达90%以上。结论完整绒毛消化联合简化的percoll密度梯度分离纯化可以得到大量较高纯度的滋养细胞以供后期实验。
[Abstract]:Objective to compare four methods for the establishment of trophoblastic cells in order to find a simple and efficient method for the culture of human trophoblastic cells in early pregnancy in vitro, and to provide a basis for related studies. Methods the villi of early pregnancy were digested and separated by trypsin and collagenase. Methods: direct inoculation, differential adhesion combined with differential digestion, isolation and purification of human peripheral blood lymphocytes, complete villus digestion combined with simplified percoll density gradient method were used to purify and culture. The morphology of cells was observed by inverted microscope, and the purity of cells was detected by HE staining, immunohistochemistry and immunocytochemistry. The positive rate of cytokeratin 7 (CK-7) expression in human peripheral blood lymphocytes was less than 60%, and the positive rate of CK-7 expression in cells obtained by simplified percoll density gradient method was more than 90%. Combined with simplified percoll density gradient separation and purification, a large number of high purity trophoblastic cells can be obtained for later experiments.
【作者单位】：承德医学院药理学教研室;承德医学院病理学教研室;承德医学院基础研究所;
【基金】：2012年河北省科学技术研究与发展计划(第五批)项目(编号:H2012406010) 河北省高等学校自然科学研究青年基金项目(编号:2010103)
【分类号】：R321

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