Primary culture and identification of mouse PASMCs and effects of hy-poxia on proliferation and apoptosis of PASMCs
10.3969/j.issn.1000-4718.2014.09.032
- VernacularTitle:小鼠肺细小动脉平滑肌细胞原代培养和鉴定以及低氧对其增殖与凋亡的影响
- Author:
Xiaoming YU
;
Rui GUO
;
Jiangfeng TANG
;
Xiaoying HUANG
;
Liangxing WANG
- Publication Type:Journal Article
- Keywords:
Pulmonary arteriolar smooth muscle Cells;
Mice;
Hypoxia;
Cell proliferation;
Apoptosis
- From:
Chinese Journal of Pathophysiology
2014;(9):1724-1728
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To establish a fast , accurate and economical technique for culturing mouse pulmonary arte-riolar smooth muscle cells ( PASMCs ) , and to explore the effects of hypoxia on the proliferation and apoptosis of the PASMCs.METHODS:In sterile condition, the pulmonary artery was isolated from the male BALB/c mice by digesting with collagenase I, and the cells were cultured in fetal bovine serum-coated flask.Centrifugal procedure was not used dur-ing the cell passage .The cell morphology was observed under an inverted phase-contrast microscope .α-Smooth muscle ac-tin was identified by immunocytochemistry and immunofluorescence .The effects of hypoxia on the proliferation and apopto-sis of the PASMCs were detected by CCK-8 assay and TUNEL assay .RESULTS:PASMCs were identified by the methods of immunocytochemistry , immunofluorescence staining and observation of morphology .Unlike the rat PASMCs with typical subcultured peak-vally pattern, the mouse PASMCs showed a lot different without a peak-vally pattern.The cells could be subcultured after 5 d to 7 d and there was 3 to 5 generations depending on the activity of the cells .CCK-8 assay demonstra-ted that the A values of PASMCs exposed to hypoxia increased after 24 h ( P<0.05) as compared with normoxia .TUNEL result showed that the apoptotic index of the PASMCs in hypoxia decreased after 24 h (P<0.05).CONCLUSION:This technique for obtaining cultured mouse PASMCs is simple , fast, accurate and economical .The digestion time is easy to control.Hypoxia promotes the proliferation and inhibits the apoptosis of PASMCs .
