Renal tubular epithelial cells are one of the essential functional cells in the kidney. Optimizing the isolation and culture method of primary renal tubular epithelial cells from SD mammary rats provides better experimental materials for renal tubule-related studies, which is essential for studying the pathogenesis of renal diseases, especially diabetic nephropathy and drug screening. SD rat renal tubular epithelial cells were isolated and purified by 2.5-mg/ml collagenase II or 2 mg/ml trypsin + 2.5 mg/ml collagenase II enzymatic digestion. The isolation and purification were observed at different time points (15 min, 30 min, 45 min, and 60 min) to determine the optimal extraction time for the enzymatic digestion method. After comparing the two enzymatic methods, it was determined that the trypsin + collagenase II enzymatic method was more effective. The primary renal tubular epithelial cells extracted by the trypsin + collagenase II digestion method were identified by the marker Cytokeratin 18 of renal tubular epithelial cells at 45 min of digestion with high purity. We established a simple, efficient, and reproducible method for isolation and culture of renal tubular epithelial cells in SD mammary gland rats.

肾小管上皮细胞是肾脏的重要功能细胞之一。优化SD乳鼠原代肾小管上皮细胞的分离培养方法，为肾小管相关研究提供更好的实验材料，对于研究肾脏疾病特别是糖尿病肾病的发病机制和药物筛选至关重要。SD大鼠肾小管上皮细胞通过2.5mg/ml胶原酶II或2mg/ml胰蛋白酶+2.5mg/ml胶原酶II酶消化分离纯化。在不同时间点（15分钟、30分钟、45分钟和60分钟）观察分离和纯化情况，以确定酶消化方法的最佳提取时间。经过两种酶解法的比较，确定胰蛋白酶+胶原酶II酶解法更为有效。采用胰蛋白酶+II型胶原酶消化法提取的原代肾小管上皮细胞，经消化45分钟后肾小管上皮细胞标记物Cytokeratin 18进行鉴定，纯度较高。我们建立了一种简单、有效、可重复的方法来分离和培养 SD 乳腺大鼠肾小管上皮细胞。