昆明小白鼠肾小管上皮细胞原代分离培养方法的建立

旨在通过比较胶原酶消化法、胰蛋白酶消化法对小白鼠肾小管上皮细胞分离效果,以及酶消化作用时间(10、20、30min)、鼠龄(成年鼠、乳鼠)对细胞产量和质量的影响研究,经细胞鉴定,建立一种小白鼠肾小管上皮细胞分离培养的方法。选用两组昆明小鼠(成年鼠和10～14日龄幼鼠),用0.1%胶原蛋白酶、0.25%的胰蛋白酶分别消化成悬液,酶的作用时间分别为10、20、30min,用倒置显微镜观察细胞培养质量,用免疫细胞化学方法鉴定细胞种类。结果表明,成年小鼠肾脏分离效果比幼鼠好;两种酶消化10min得到的细胞产量最多;胶原酶消化法比胰蛋白酶消化法分离的细胞贴壁早,且贴壁数量多;细胞鉴定结果为阳性。结论:对成年昆明小鼠肾脏,用胶原酶消化法,消化10min,肾小管上皮细胞产量和质量较好。

The establishment of primary renal tubular epithelial cells isolated and culture method from Kumming Mus musculus albus

(Objective) To through comparing the isolation yield and quality of renal tubular epithelial cells by two different digestion methods (collagenase and trypsinase digestion method), different digested time (10、20、30 min) , in rats in two different ages (suckling and adult), and then verificating the category of the cells ,to establish the Method ofisolatingand culturingprimarytubular epithelial cell fromprimary tubular epithelial cell. (Methods) The tubular epithelial cells get from suckling and adult Kumming Mus musculus albus was digested by two different methods, respectively, after digestion of 10min,20min and 30min;then observe the qualityofthe cells with inverted microscope and verificate them by immuocytochemistry Method. The results show that , The yield of primary tubular epithelial cells by adult group were better than suckling group;the two enzyme digestion methods were suitable in 10 min’digestion. The collagenase digestion method is better than trypsinase digestion method in cells adherence rate and quantity; identity result is positive. (Conclusion)The primary tubular epithelial cell is better get from the kidney of adult KunmingMus musculus albus ,which is better digested 10min bycollagenase.