摘要
目的:分离大鼠胎鼠小肠黏膜来源的边缘群(SP)细胞,并分析其生物学特性。方法:取Wistar大鼠胎鼠小肠全长,按文献[3]方法制作小肠黏膜的类器官片段并制备成单细胞悬液。使用Hoechst33342和PI染色后用流式细胞仪分选SP细胞。提取分选前后的细胞总RNA和蛋白,用实时定量PCR及Western-blot方法分别检测其中MSI-1RNA及MSI-1蛋白的表达水平。结果:大鼠胎鼠来源的小肠黏膜单细胞悬液中包含一个特定的细胞群体—SP细胞,染色液中加入维拉帕米后,SP细胞被阻断后消失。定量PCR检测显示SP细胞中MSI-1mRNA水平是分选前的8.125倍,是非SP细胞的20.077倍。Western-blot结果显示MSI-1蛋白水平明显高于分选前。结论:大鼠胎鼠的小肠黏膜SP细胞富集小肠黏膜干细胞,与小鼠来源SP细胞生物学特性基本一致。
Objective ：To isolate side population（SP） cells from fetal intestine Wistar rat and to analyze their properties. Methods ： Intestine organoids of fetal rat were isolated and dissociated into single cells. These cells were stained with Hoechst 33342 and propidium iodide and then sorted using fluorescence-activated cell sorting. Total RNA and protein were purificated from sorted fractions to detect Musashi-1 expression by real-time reverse-transcription polymerase chain reaction and Westem-bloting. Results ： Fetal Wistar rat intestine contained a viable population of cells that showed the side population phenotype and were sensitive to verapamil. These cells were 8. 125-fold enriched for Musashi-1 mRNA compared with MSI-1 protein. thelial cells is intact tissue and 20 Conclusion ： The sid enriched for intestinal fraction derived from mice 7-fold compared with non-SP cells. They were also enriched for e population fraction separated from fetal Wistar rat intestinal epistem ceils, which has similar characteristics to side population
出处
《医学研究生学报》
CAS
2007年第7期693-696,共4页
Journal of Medical Postgraduates
基金
国家自然科学基金资助项目(批准号:30125018)
中国博士后科学基金资助项目(批准号:2006040938)
关键词
干细胞
边缘群细胞
小肠黏膜
Stem cells
Side population cells
Intestinal mucosa
