CAJ | 학술논문

目的建立较好的大鼠近端肾小管上皮细胞原代培养模型.方法无菌取Wistar大鼠肾脏,取皮质剪碎,经Ⅰ型胶原酶消化和45%Percoll连续密度梯度离心进行纯化,用含10%胎牛血清的DMEM/F12培养基原代培养并传代,观察细胞形态并以免疫细胞化学染色及酶化学染色鉴定.结果培养4～5 d细胞融合成单层,呈典型的鹅卵石样,细胞角蛋白18及碱性磷酸酶染色均呈阳性,细胞可传2～3代.结论此法可在短期获得数量较多、重复性好的近端肾小管上皮细胞,为体外研究肾小管细胞的病变提供了实验平台.
목적 건립교호적대서근단신소관상피세포원대배양모형.방법 무균취Wistar대서신장,취피질전쇄,경Ⅰ형효원매소화화45%Percoll련속밀도제도리심진행순화,용함10%태우혈청적DMEM/F12배양기원대배양병전대,관찰세포형태병이면역세포화학염색급매화학염색감정.결과 배양4～5 d세포융합성단층,정전형적아란석양,세포각단백18급감성린산매염색균정양성,세포가전2～3대.결론 차법가재단기획득수량교다、중복성호적근단신소관상피세포,위체외연구신소관세포적병변제공료실험평태.
Objective To establish the better model for primary culture of rat proximal tubule epithelial cells(PTCs).Methods Wistar rats kidney were sterile taken out, then the cortex was separated and shredded, purified by collagenase Ⅰdigestion and 45% percoll discontinuous density gradient centrifugation, cultured and passaged by DMEM/F12 medium supplemented with 10% fetal bovine serum, identified by morphology, immunocytochemistry staining and enzyme staining.Results After 4 to 5 days,the cells were inosculated into a single layer and showed the typical cobblestone-like appearance.The cytokeratin18 and alkaline phosphatase staining were positive, the cells could be passed 2～3 generation.Conclusions This method can yield large quantity and good repeatability PTCs in short term,those cells can provide an experimental platform for the research of pathological changes and lesions.