摘要
目的:建立大鼠和小鼠胃平滑肌细胞的分离、鉴定、原代培养及评价方法。方法:取SD大鼠和C57BL/6小鼠各30只,分别随机分为3组,每组10只,进行3次平行实验,无菌条件下分离原代胃平滑肌细胞。采用形态学观察、免疫荧光与蛋白印迹等方法对分离的细胞进行鉴定,流式细胞术分析细胞纯度,台盼蓝拒染法评价复苏后的细胞活力。结果:分离的大鼠和小鼠原代胃平滑肌细胞呈梭形或多边形;免疫荧光与蛋白印迹结果显示大鼠和小鼠胃平滑肌细胞均能表达α-平滑肌肌动蛋白;流式细胞术结果显示大鼠胃平滑肌细胞中阳性表达率为99.6%,小鼠胃平滑肌细胞中阳性表达率为97.4%。冻存后复苏的大鼠和小鼠胃平滑肌细胞的活细胞率可以达到97%以上。结论:本研究建立了稳定、可靠的大鼠和小鼠原代胃平滑肌细胞分离培养方法与评价指标。
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 ofα-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate ofα-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.
作者
黄佳怡
陈琼
杨子龙
宗阳
张露蓉
王斐
袁琴
HUANG Jiayi;CHEN Qiong;YANG Zilong;ZONG Yang;ZHANG Lurong;WANG Fei;YUAN Qin(Central Laboratory,Suzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine,Clinical Pharmaceutical Laboratory of Traditional Chinese Medicine,Suzhou Academy of Wumen Chinese Medicine,Suzhou215009,China)
出处
《中国病理生理杂志》
北大核心
2025年第8期1658-1664,共7页
Chinese Journal of Pathophysiology
基金
国家自然基金青年科学基金项目(No.81903899)
苏州市科技发展计划项目(No.SKY2023217)。
关键词
胃平滑肌细胞
鉴定
Α-平滑肌肌动蛋白
原代培养
流式细胞术
gastric smooth muscle cells
identification
α-smooth muscle actin
primary culture
flow cytometry
