CAJ | 학술논문

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.
목적 탐토밀도제도리심법작위일충신형장상피간세포분리방법적가행성.방법 통과복강주사대제량5-불뇨밀정(5-FU)제비장점막엄중손상소서모형,취출손상적장점막소화성단세포현액,이용Percoll밀도제도리심법분리세포체적대소불동적세포군.HE염색관찰각군세포형태특정,면역세포화학급RT-PCR검측각군세포musashi-1(msi-1)적표체.결과 분리적대부분세포위우50%화70%밀도제도적Percoll분리액체층,50%밀도제도분리액체층중적세포부합간세포적형태특정,면역세포화학검측기msi-1표체양성솔약위93%,RT-PCR현시해군세포msi-1 mRNA표체교강.결론 건립료일충간편、유효적분리구유활성적장상피간세포적방법.
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.