基于细胞骨架差异的心肌细胞纯度快速鉴定方法  

Rapid Identification of Myocardial Cell Purity Based on Cytoskeletal Differences

作　　者：潘丛彬 王思琪 赵宝清 尧青[2] 郭西英 欧阳昌汉[1] 任展宏 刘超[2] PAN Cong-bing;REN Zhan-hong;LIU Chao(School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning Hubei 437100,China)

机构地区：[1]湖北科技学院医学部药学院,湖北咸宁437100 [2]湖北科技学院糖尿病心脑血管病变湖北省重点实验室,湖北咸宁437100

出　　处：《湖北科技学院学报（医学版）》2023年第2期104-109,F0002,共7页Journal of Hubei University of Science and Technology(Medical Sciences)

基　　金：国家自然科学基金面上项目(81870173,82073852);湖北省自然科学基金青年项目(2022CFB843);湖北省重点实验室糖尿病专项基金项目(2022TNB04);湖北省教育厅中青年人才项目(Q20202803);咸宁市科技计划自然科学基金项目(2021ZRKX024);湖北科技学院博士启动基金项目(BK202121;BK202027);湖北科技学院科研创新团队项目(2022T01)。

摘　　要：目的寻找并确定一种优于传统且简单快速的心肌细胞鉴定方法。方法取1~3d龄SD大鼠心脏,0.1%Ⅱ型胶原酶消化,差速贴壁分离心肌细胞与心肌成纤维细胞,对心肌细胞使用5-溴脱氧尿嘧啶进行纯化,并通过传代对心肌成纤维细胞进行纯化。采用免疫荧光法,Phalloidin固定染色和Tubulin Tracker活细胞染色鉴定心肌细胞与心肌成纤维细胞。结果心肌细胞的微丝与微管可分别被Phalloidin和Tubulin Tracker标记,而心肌成纤维细胞的微丝和微管均未被标记,且这两种染色方法耗时极少。结论基于细胞骨架差异的特征,通过标记微丝或者微管能够简便快速鉴别心肌细胞和心肌成纤维细胞。本方法明显优于免疫荧光染色,将提高心血管疾病在细胞水平上的研究效率。Objective To find and determine a simple and rapid identification method for cardiomyocytes,which is better than the traditional method.Methods Heart tissues isolated from 1~3 days SD neonatal rats were digested with 0.1%collagenase typeⅡ.Then,cardiomyocytes and myocardial fibroblasts were isolated by differential attachment.The cardiomyocytes were furtherly purified under 5-bromodeoxyuracil treatment while myocardial fibroblasts were purified by cell cultures.Cardiomyocytes and myocardial fibroblasts were identified by immunofluorescence staining,phalloidin staining and Tubulin Tracker live cell staining.Results The microfilaments and microtubules of cardiomyocytes could be stained with phalloidin and Tubulin Tracker,respectively,while the those of myocardial fibroblasts were not stained.Conclusion Cardiomyocytes and myocardial fibroblasts can be easily and rapidly identified by staining microfilaments or microtubules due to their cytoskeleton distinction.This method is obviously superior to the immunofluorescence staining,and furtherly improve the research efficiency of cardiovascular disease at the cell level.

关 键 词：心肌细胞 心肌成纤维细胞 鬼笔环肽 微丝 微管 细胞鉴定 

分 类 号：R-332[医药卫生]