小鼠小肠上皮细胞的体外原代培养

目的探讨小鼠小肠上皮细胞原代培养的原理和方法。方法取胎龄为17~19d的昆明鼠的小肠,通过3种不同的小肠上皮细胞分离方法进行原代培养。结果体外培养出圆形、多角形单层生长,呈"铺路石样",核周可见许多小空泡并且CK18、碱性磷酸酶染色阳性的细胞。结论应用中性蛋白酶Ⅰ和粗胶原酶Ⅺ消化可获满意的上皮细胞分离效果,细胞贴壁良好。小肠上皮在体外的增殖分化有赖于分离的细胞器中干细胞及短暂分裂细胞的多少以及一些间质细胞的存在。

In Vitro Primary Culture of Mouse Intestinal Epithelial Cells

Objective To explore the principle and method of primary culture of mouse intestinal epithelial cells.Methods The intestinal epithelial cells were taken from embryo 17-19 d mouse for primary culture by three different methods.Results These cells of primary culture in vitro were round or polygonal cells,grew into pavestone-like monolayer,had many small vacuoles around nucleus,and were the positive of cytokeratin 18 and Alkaline phosphatase staining.Conclusion The intestinal epithelial cells in better condition can be obtained by dispase I and crude collagenase XI digestion in primary culture in vitro.Proliferation of the intestinal epithelial cells in vitro was dependent upon the number of the intestinal stem cells and transient amplifying cells.As also,mesenchymal cells were important factors to proliferation of the intestinal epithelial cells.