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Primary culture and identification of mouse renal proximal tubular epithelial cells

WANG Li-Ming, CHEN Jia, CHEN Ke-Hong, CAI Ming-Yu, WANG Xiao-Yue, HE Ya-Ni^*

Department of Nephrology, Daping Hospital, Army Military Medical University, Chongqing 400042, China

Abstract

The present study was aimed to establish a modified method for culturing mouse renal proximal tubular epithelial cells (TECs). Renal cortex was isolated from mouse kidney and scissored into pieces. TECs were separated by digesting scissored renal cortex in type II collagenase combined with strainer filtration, and then cultured in DMEM. The morphology of TECs was observed under inverted microscopy. The cell proliferative ability was assessed by flow cytometry, and cell viability was analyzed by CCK-8 assay. The purity of TECs was identified by immunofluorescence. Immunofluorescence observation showed that more than 95% cells were epithelial marker CK18 positive and more than 90% cells expressed renal proximal TECs marker proteins, Villin, AQP1, and SGLT2. The cells could be subcultured for about 5 times. The cell proliferative ability declined following the repeated passage. This study introduced a modified efficient method for culturing highly purified mouse renal proximal TECs.

Key words: renal tubular epithelial cells; primary cell culture; mouse

Received: 2017-12-07　　Accepted: 2017-12-07

Corresponding author: 何娅妮　　E-mail: Heynmail@163.com

DOI: 10.13294/j.aps.2018.0046

Citing This Article：

WANG Li-Ming, CHEN Jia, CHEN Ke-Hong, CAI Ming-Yu, WANG Xiao-Yue, HE Ya-Ni. Primary culture and identification of mouse renal proximal tubular epithelial cells. Acta Physiol Sin 2018; 70 (4): 406-412 (in Chinese with English abstract).