Establishment and functional characterization of the reversibly immortalized mouse glomerular podocytes(imPODs)  

作　　者：Xinyi Yu Liqun Chen Ke Wu Shujuan Yan Ruyi Zhang Chen Zhao Zongyue Zeng Yi Shu Shifeng Huang Jiayan Lei Xiaojuan Ji Chengfu Yuan Linghuan Zhang Yixiao Feng Wei Liu Bo Huang Bo Zhang Wenping Luo Xi Wang Bo Liu Rex C.Haydon Hue H.Luu Tong-Chuan He Hua Gan 

机构地区：[1]Departments of Nephrology,Orthopaedic Surgery,Cardiology,General Surgery,Plastic Surgery,and Clinical Laboratory Medicine,The First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China [2]Molecular Oncology Laboratory,Department of Orthopaedic Surgery and Rehabilitation Medicine,The University of Chicago Medical Center,Chicago,IL,60637,USA [3]Ministry of Education Key Laboratory of Diagnostic Medicine and School of Laboratory Medicine,and the Affiliated Hospitals of Chongqing Medical University,Chongqing,400016,China [4]Department of Biochemistry and Molecular Biology,China Three Gorges University School of Medicine,Yichang,443002,China [5]Department of Clinical Laboratory Medicine,The Second Affiliated Hospital of Nanchang University,Nanchang,330031,China [6]Key Laboratory of Orthopaedic Surgery of Gansu Province and the Department of Orthopaedic Surgery,The Second Hospital of Lanzhou University,Lanzhou,730030,China

出　　处：《Genes & Diseases》2018年第2期137-149,共13页基因与疾病（英文）

基　　金：The reported work was supported in part by research grants from the National Institutes of Health(CA226303 to TCH);the National Key Research and Development Program of China(2016YFC1000803 and 2011CB707906 to TCH);This project was also supported in part by The University of Chicago Cancer Center Support Grant(P30CA014599);the National Center for Advancing Translational Sciences of the National Institutes of Health through Grant Number UL1 TR000430.

摘　　要：Glomerular podocytes are highly specialized epithelial cells and play an essential role in establishing the selective permeability of the glomerular filtration barrier of kidney.Maintaining the viability and structural integrity of podocytes is critical to the clinical management of glomerular diseases,which requires a thorough understanding of podocyte cell biology.As mature podocytes lose proliferative capacity,a conditionally SV40 mutant tsA58-immortalized mouse podocyte line(designated as tsPC)was established from the Immortomouse over 20 years ago.However,the utility of the tsPC cells is hampered by the practical inconvenience of culturing these cells.In this study,we establish a user-friendly and reversibly-immortalized mouse podocyte line(designated as imPOD),on the basis of the tsPC cells by stably expressing the wildtype SV40 T-antigen,which is flanked with FRT sites.We show the imPOD cells exhibit long-term high proliferative activity,which can be effectively reversed by FLP recombinase.The imPOD cells express most podocyte-related markers,including WT-1,Nephrin,Tubulin and Vinculin,but not differentiation marker Synaptopodin.The imPOD cells do not form tumor-like masses in vivo.We further demonstrate that TGFb1 induces a podocyte injury-like response in the FLP-reverted imPOD cells by suppressing the expression of slit diaphragm-associated proteins P-Cadherin and ZO-1 and upregulating the expression of mesenchymal markers,a-SMA,Vimentin and Nestin,as well as fibrogenic factors CTGF and Col1a1.Collectively,our results strongly demonstrate that the newly engineered im-POD cells should be a valuable tool to study podocyte biology both under normal and under pathological conditions.

关 键 词：Chronic kidney disease FLP recombinase Glomerular disease GLOMERULUS IMMORTALIZATION NEPHROPATHY PODOCYTE SV40 T antigen 

分 类 号：R69[医药卫生—泌尿科学]