畜牧兽医学报2017，Vol.48Issue(1)：75-82,8.DOI:10.11843/j.issn.0366-6964.2017.01.009
猪胎盘绒毛滋养层细胞体外原代培养
Primary Culture of Villous Cytotrophoblasts Isolated from Pig Fullterm Placenta
摘要
Abstract
This research was conducted to establish a simple and effective method for isolation and purification of pig chorionic trophoblast cells.Single cell suspension was prepared from pig fullterm placentas by the method of trypsin-DNase Ⅰ digestion and two density (35％ and 45％,v/v)Percoll gradient centrifugation.Morphology observation,growth curve measurement,immunofluorescence and transmission electron microscopy (TEM) were applied to cells identification.The results showed that the cells were polygon or quasi-circular in shape,which exhibited epithelioid and paving-like spreading growth.After 48 hours culture,some cells started developing into syncytiotrophoblasts.Immunofluorescence assay showed that the proportion of Cytokeratin 7 positive cells was more than 91 ％,indicated the high purity of pig chorionic trophoblast cells.The results of TEM revealed that the cells had typical structural features of trophoblasts.These results indicated that our research could be used to obtain high-purity pig chorionic trophoblast cells simply and effectively by the method of trypsin-DNase Ⅰ digestion and 2 density (35％ and 45％,v/v) Percoll gradient centrifugation.关键词
猪/胎盘/滋养层细胞/原代培养Key words
pig/placenta/villous cytotrophoblasts/primary culture分类
农业科技引用本文复制引用
董书圣,田亮,颜培实..猪胎盘绒毛滋养层细胞体外原代培养[J].畜牧兽医学报,2017,48(1):75-82,8.基金项目
江苏省自然科学基金-青年基金项目(BK20150672) （BK20150672）
