改良的原代心肌细胞培养方法
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- 英文论文题名：An Improved Method for Primary Culture of Cardiomyocytes
- 论文作者：赵勇 ; 徐振宇 ; 吕少春 ; 赵隽鹏 ; 李梅秀 ; 刘晓伟 ; 田国忠
- 英文论文作者：ZHAO Yong ; XU Zhen-yu ; LV Shao-chun ; ZHAO Jun-peng ; LIU Xiao-wei ; LI Mei-xiu ; TIAN Guo-zhong ; Basic Medical College of Jiamusi University
- 年：2017
- 作者机构：佳木斯大学基础医学院;
- 论文关键词：心肌细胞 ; 原代培养 ; 新生大鼠
- 英文论文关键词：Cardiomyocytes ; Primary culture ; Neonatal rat
- 会议召开时间：2017-08-07
- 会议录名称：中国解剖学会第六届全国解剖学技术学术会议论文汇编
- 语种：中文
- 分类号：Q813.11
- 学会代码：ZGJP
- 会议名称：中国解剖学会第六届全国解剖学技术学术会议
- 会议地点：中国江西九江
- 主办单位：中国解剖学会科技开发和咨询工作委员会
- 学会名称：中国解剖学会
- 页数：2
- 文件大小：1942k
- 原文格式：D
- 会议级别：全国
摘要
目的探讨乳鼠原代心肌细胞的培养方法,简便稳定地获得高存活率和纯度的心肌细胞。方法无菌条件下取3日内新生SD大鼠的心室肌组织,首先用胰蛋白酶消化心肌组织一次,再用I型胶原酶短时多次消化,然后应用差速贴壁分离法和化学试剂抑制法纯化心肌细胞。对心肌细胞进行形态学观察、存活率检测和免疫荧光鉴定。结果台盼蓝染色检测心肌细胞存活率在95%以上。培养24小时后的心肌细胞已贴壁生长,呈梭形及多边形,有自发性搏动,48小时后细胞伪足相互交织成网,进而形成细胞簇,呈现同步性搏动。免疫荧光鉴定心肌细胞纯度达96%。结论此种方法能够可靠地获得较高存活率和纯度的原代乳鼠心肌细胞。
Objective To explore the culture method of primary cardiomyocytes in neonatal rats,and to obtain high survival rate and purity cardiomyocytes.Methods Under aseptic condition,the ventricular muscle tissue of neonatal SD rats(born within 3 days) was obtained.First of all,the myocardial tissue was digested with trypsin,and then the type I collagenase was used for several times.Then,purified by differential adhesion and chemical reagent inhibition method.Morphological observation,survival rate and immunofluorescence identification of cardiomyocytes were performed.Results Trypan blue staining was used to detect the survival rate of myocardial cells above 95%.After 24 hours of culture,the myocardial cells have been adherent growth,which were spindle shaped and polygonal,with spontaneous pulsation.After the first 48 hours,the cells were intertwined with each other to form a cluster of cells,showing synchronous pulse.The purity of myocardium was 96% by immunofluorescence.Conclusion This method can reliably obtain a higher survival rate and purity of primary neonatal rat cardiomyocytes.
Objective To explore the culture method of primary cardiomyocytes in neonatal rats,and to obtain high survival rate and purity cardiomyocytes.Methods Under aseptic condition,the ventricular muscle tissue of neonatal SD rats(born within 3 days) was obtained.First of all,the myocardial tissue was digested with trypsin,and then the type I collagenase was used for several times.Then,purified by differential adhesion and chemical reagent inhibition method.Morphological observation,survival rate and immunofluorescence identification of cardiomyocytes were performed.Results Trypan blue staining was used to detect the survival rate of myocardial cells above 95%.After 24 hours of culture,the myocardial cells have been adherent growth,which were spindle shaped and polygonal,with spontaneous pulsation.After the first 48 hours,the cells were intertwined with each other to form a cluster of cells,showing synchronous pulse.The purity of myocardium was 96% by immunofluorescence.Conclusion This method can reliably obtain a higher survival rate and purity of primary neonatal rat cardiomyocytes.
引文
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[2]王涛,余志斌,谢满江,等.新生大鼠心肌细胞培养技巧[J].第四军医大学学报,2003,24(2):1.
[3]Louch WE,Sheehan KA,Wolska BM.Methods in cardiomyocyte enisolation,culture,and gene transfer[J].J Mol Cell Cardiol,2011,51(3):288-298.
[4]张晓京,张建东,来丽娜,等.SD乳鼠原代心肌细胞培养方法的改进[J].长治医学院学报,2012,3(26):171-173.
[2]王涛,余志斌,谢满江,等.新生大鼠心肌细胞培养技巧[J].第四军医大学学报,2003,24(2):1.
[3]Louch WE,Sheehan KA,Wolska BM.Methods in cardiomyocyte enisolation,culture,and gene transfer[J].J Mol Cell Cardiol,2011,51(3):288-298.
[4]张晓京,张建东,来丽娜,等.SD乳鼠原代心肌细胞培养方法的改进[J].长治医学院学报,2012,3(26):171-173.