摘要
目的:建立一种经济、高效的人足月胎盘滋养细胞分离培养方法。方法:收取正常足月剖宫产患者胎盘组织,DNA酶联合胰酶反复消化、网筛过滤、不连续密度梯度分离液分离纯化细胞;免疫细胞化学法鉴定细胞来源;对所培养的滋养细胞进行形态学观察;CCK-8增殖实验分析细胞体外增殖活力。结果:利用此法可成功体外培养胎盘滋养细胞,一次操作可获得大于107个细胞。体外培养时,滋养细胞增殖能力低下,3h开始贴壁,24h后逐渐开始融合成合体滋养细胞。结论:本研究改进了既往足月胎盘滋养细胞原代培养的方法,简化操作步骤,缩短操作时间,并且一次性可获得大量且纯度较好的胎盘滋养细胞。
Objective： To establish an economical and efficient method for isolating, purificating and culturing term human placental trophoblasts. Methods. Human placenta of normal pregnancy was obtained right after cesarean section, the tissue was digested repeatedly with Deoxyribonuclease and trypsion, mesh screen filration and discontinuous density gradient centrifugation were used to isolate and purify the trophblast cells. Morphological changes of cells were observed under microscope and identified by immunohistochemical staining. CCK-8 proliferation assay was employed to analyze the viability of trophblats in vitro. Results： Trophbalst cells could be isolated and cultured successfully in vitro with this method. More than 107 cells can be obtained at a time from 30 to 40 g tissues, the proliferation capacity of trophbalst cells is low in vitro. Cells in vitro began to stick wall after 3 hours and gradually began to merge into syncytiotrophoblasts after 24 hours. Conclusion： This study improves the isolating and culturing method of term human placental trophoblasts, and huge amount of purified cells can be obtained with more simplified operation in a rather short period of time.
出处
《武汉大学学报（医学版）》
CAS
2015年第5期813-816,共4页
Medical Journal of Wuhan University
基金
国家自然科学基金面上项目(编号:81370707
81270753)
关键词
人足月胎盘
滋养细胞
原代培养
Term Human Placent
Trophblast Cells
Primary Culture
