小鼠肾小管上皮细胞的原代培养及鉴定

目的 建立小鼠肾小管上皮细胞(mRTECs)原代培养、传代及鉴定的方法.方法 采用肾小管节段贴块培养法与3种消化培养法(胰蛋白酶、胰蛋白酶 EDTA、胶原酶Ⅰ)进行小鼠肾小管上皮细胞原代培养,锥虫蓝染色后镜下观察消化后细胞成活率、细胞形态和数量.胰蛋白酶消化传代,以免疫细胞化学方法鉴定细胞种类.结果 贴块法和胶原酶Ⅰ消化培养法均能成功培养小鼠肾小管上皮细胞,两者相比,胶原酶Ⅰ消化培养法培养的细胞生长更快.结论 胶原酶Ⅰ消化培养法是小鼠肾小管上皮细胞原代培养的理想方法.

PRIMARY CULTURE AND IDENTIFICATION OF MOUSE RENAL TUBULAR EPITHELIAL CELLS

Objective To establish a method for the primary culture, subculture and identification of mouse renal tubular epithelial cells(mRTECs). Methods Renal tubular segments sticking and three kinds of digesting(trypsin,trypsin+EDTA, collagenase Ⅰ) were used in the primary culture of mRTECs respectively.Cell vitality, morphology and quantity were observed and evaluated by phase-contrast microscopy after trypan-blue staining. Trypsin digestion was used in the subculture of mRTECs. The types of cells were identified by cellular immunocytochemistry. Results The culturing methods of renal tubular segments sticking and collagenase Ⅰ digesting were successfully used in the primary culture of mRTECs and the latter was quicker for the growth of cells.Conclusion Collagenase Ⅰ digesting is an ideal method of cellular culture for the primary culture of mouse renal tubular epith