一种新型肠上皮干细胞分离方法的建立  

Establishment of a novel method for isolation of intestinal epithelium stem cells

作　　者：徐波[1] 罗育其 肖焕擎[1] 蔡文松[1] 于海涛 董栋 贾林[2] 

机构地区：[1]广州医学院附属广州市第一人民医院普外科,510180 [2]广州医学院附属广州市第一人民医院消化内科,510180 [3]广州市南沙区中心医院外科

出　　处：《中华生物医学工程杂志》2011年第2期139-142,共4页Chinese Journal of Biomedical Engineering

基　　金：广州市医药卫生科技项目（2009-YB-016）

摘　　要：目的 探讨密度梯度离心法作为一种新型肠上皮干细胞分离方法的可行性.方法 通过腹腔注射大剂量5-氟尿嘧啶（5-FU）制备肠黏膜严重损伤小鼠模型,取出损伤的肠黏膜消化成单细胞悬液,利用Percoll密度梯度离心法分离细胞体积大小不同的细胞群.HE染色观察各群细胞形态特征,免疫细胞化学及RT-PCR检测各群细胞musashi-1（msi-1）的表达.结果 分离的大部分细胞位于50%和70%密度梯度的Percoll分离液体层,50%密度梯度分离液体层中的细胞符合干细胞的形态特征,免疫细胞化学检测其msi-1表达阳性率约为93%,RT-PCR显示该群细胞msi-1 mRNA表达较强.结论 建立了一种简便、有效的分离具有活性的肠上皮干细胞的方法.Objective To validate the establishment of a novel method for isolation of intestinal epithelium stem cells using density gradient centrifugation.Methods The model of severe small intestinal mucosa damage in mice was established by intraperitoneal injection of high-dose 5-fluorouracil（5-FU）. Then the damaged intestinal mucosa was digested and prepared into single cell suspension.Percoll gradient centrifugation was used to separate the cells into subsets based on their sizes. The morphological characteristics of these cell subsets were studied with HE staining, and the musashi-1 expression was detected by immunocytochemistry and RT-PCR.Results Most of the cells were located in the 50% and 70% layers of Percoll separating medium.The cells in the 50% layer displayed stem cell-like morphology. Immunocytochemistry showed positive musashi-1 expression in about 93% of these cells, and RT-PCR also revealed high level of musashi -1 expression.Conclusion A simple and effective method for isolation of viable intestinal stem cells was established in this study.

关 键 词：离心法 梯密度 干细胞 肠黏膜 细胞分离 实验室技术和方法 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]