Isolation,identification,and primary culture of gastric smooth muscle cells derived from rats and mice
10.3969/j.issn.1000-4718.2025.08.025
- VernacularTitle:大鼠和小鼠胃平滑肌细胞的分离、鉴定及原代培养
- Author:
Jiayi HUANG
1
;
Qiong CHEN
1
;
Zilong YANG
1
;
Yang ZONG
1
;
Lurong ZHANG
1
;
Fei WANG
1
;
Qin YUAN
1
Author Information
1. 南京中医药大学附属苏州市中医医院中心实验室,苏州市吴门医派研究院中药临床药学实验室,江苏 苏州 215009
- Publication Type:Journal Article
- Keywords:
gastric smooth muscle cells;
identification;
α-smooth muscle actin;
primary culture;
flow cytometry
- From:
Chinese Journal of Pathophysiology
2025;41(8):1658-1664
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.
