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目的 :探讨大鼠小肠上皮细胞原代培养的原理与方法以及培养过程中常见的问题和克服办法。方法 :取生后 1周SD大鼠的小肠上皮细胞 ,经分离纯化后进行原代培养。结果与结论 :胶原酶Ⅺ型和中性蛋白酶Ⅰ型配合使用 ,可以分离到形态完整、容易贴壁生长的小肠绒毛细胞团。进一步利用离心技术纯化得到 95 %以上的小肠上皮细胞。结果显示 ,常规体外培养的肠上皮细胞是否增殖分化最终决定于用酶消化后的结构完整性 ,如果用0 .2 5 %胰酶消化 ,则细胞受损程度较大 ,不易贴壁生长。
Objective: To explore the principle and method of primary culture of rat small intestinal epithelial cells and the common problems and solutions in the process of culture. Methods: The small intestine epithelial cells of SD rats were isolated and purified after 1 week. RESULTS AND CONCLUSION: Collagenase Ⅺ type and neutral protease type Ⅰ could be used to isolate small intestine villous cell mass with complete morphology and easily adherent growth. Further use of centrifugal technology to obtain more than 95% of intestinal epithelial cells. The results showed that whether the proliferation and differentiation of intestinal epithelial cells cultured in vitro is ultimately determined by the structural integrity after digestion with enzymes, if 0.2% trypsin digestion, the cell damage is greater, difficult to adherent growth.