Culture and identification of parietal epithelial cells of Bowman′s capsule
10.3760/cma.j.cn441217-20211206-00074
- VernacularTitle:鲍曼囊壁层上皮细胞培养与鉴定
- Author:
Minzhou WANG
1
;
Chenqi XU
;
Lulin MIN
;
Jiayue LU
;
Xiajing CHE
;
Renhua LU
;
Kewei XIE
;
Leyi GU
Author Information
1. 上海交通大学医学院附属仁济医院肾脏科 分子细胞肾病实验室，上海 200127
- Keywords:
Nephrons;
Epithelial cells;
Cell separation;
Cells, cultured;
Identification
- From:
Chinese Journal of Nephrology
2022;38(6):536-542
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the isolation and culture methods of mouse parietal epithelial cells (PECs) of Bowman′s capsule, so as to provide a cell tool for further study.Methods:Mouse renal corpuscles were isolated by cell sieving combined with magnetic separation. After primary culture, identified parietal epithelial cells were induced to differentiate into podocytes. Immunofluorescence staining, real-time quantitative PCR and Western blotting were used to detect specific markers of parietal epithelial cells and podocytes.Results:Primary cultured PECs grew like paving stone and expressed Claudin-1 (PECs specific marker), CD133 (stem cell marker) and CD24 (stem cell marker), without the expression of tubular epithelial cell proteins, mesangial cell and podocyte specific proteins. Cultured to 6 generations in vitro, the PECs still expressed Claudin-1, CD133 and CD24. After incubated with differentiation medium, PECs were able to express podocyte markers WT-1 and Synaptopodin. Conclusion:The renal corpuscles are extracted by cell sieving combined with magnetic separation, and the mouse PECs successfully cultured in vitro can be induced to express podocytes′ markers.
