Primary culture and identification of rat glomerular microvascular endothelial cells.
- Author:
Hua-Gen MA
1
;
Hai-Qin LIU
2
;
Zhao-De LIU
3
;
Yuan-Yu TANG
4
Author Information
1. Department of Shang Han, College of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China.
2. College of Integrated Traditional Chinese and Western Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China.
3. Basic Medical College, Nanjing Medical University, Nanjing 211116, China.
4. College of Traditional Chinese Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China. 2422198977@qq.com.
- Publication Type:Journal Article
- MeSH:
Animals;
Cells, Cultured;
Endothelial Cells;
Rats;
Rats, Sprague-Dawley
- From:
Acta Physiologica Sinica
2021;73(6):926-930
- CountryChina
- Language:Chinese
-
Abstract:
The aim of the present study was to establish a simple and efficient method for isolation and culture of primary rat glomerular microvascular endothelial cells in vitro. The bilateral kidneys were taken from 7-10-day old Sprague-Dawley rats, and the renal cortex was separated. Glomeruli were obtained by cutting and continuously passing 200-mesh and 300-mesh sieves. After type IV collagenase digestion for 15-20 min, renal microvascular globules were collected for inoculation and culture. The cultured cells were identified by cell morphology observation and immunocytochemical staining with factor VIII related antigen. The results showed that the renal microvascular globules were irregularly spherical, without cysts, and the capillary loop structure was clear; after 3 days of primary culture, short spindle-shaped cells crawled out around the renal microvascular globules and gradually formed cell colonies, showing an "island-like shape" distribution; 4-5 days later, the cell colonies fused with each other; 6 days later, the cells covered the bottom of the dish, showing a typical monolayer, paving stone-like, mosaic arrangement. The immunocytochemical staining of factor VIII related antigen showed that the cytoplasm was lightly stained brownish red, and factor VIII related antigen-positive rate of cells was nearly 100%. The above results suggested that this study successfully established a method combining continuous screening and collagenase digestion for culture of primary rat glomerular microvascular endothelial cells in vitro. It provides an important tool cell for studying the mechanism of the occurrence and development of diabetic nephropathy.
