乳小鼠心肌细胞体外培养方法的优化  

An Improved Protocol for Primary Culture of Cardiomyocyte from Neonatal Mice

作　　者：张晶[1] 朱劲舟[2] 张瑞岩[2] 王霞[3] 

机构地区：[1]上海医药集团中央研究院,201203 [2]上海交通大学医学院附属瑞金医院心血管内科,200025 [3]上海交通大学药学院再生医学实验室,200240

出　　处：《中国医疗前沿》2012年第10期8-9,15,共3页China Healthcare Innovation

摘　　要：目的探讨乳小鼠心肌细胞分离和体外培养的方法,并结合实际情况建立一种优化的体外培养乳小鼠心肌细胞的方法。方法胰酶-Ⅱ型胶原酶混合液分次消化乳小鼠心肌组织后行差速贴壁法达到心肌细胞纯化目的,并利用心肌细胞特意表达a-肌动蛋白的特性运用细胞免疫组织化学方法鉴定心肌细胞。结果差速贴壁48h后心肌细胞贴壁完全,并见大部分贴壁细胞出现搏动,但未观察到同簇搏动,经a-肌动蛋白细胞免疫组织化学鉴定,心肌细胞纯度达96.7%。结论本研究优化了原代乳小鼠心肌细胞分离和体外培养方法,为后续细胞实验打下了基础。Objective To find an improved method for isolation and cultivation of cardiomyocytes from neonatal mice based on the present protocol.Methods The hearts were digested by trypsin and collagenase typeⅡ.The cell suspension was preplated to reduce the fibroblast content.Immunocytochemistry with primary antibody of a-actin was used to demonstrate that cultured cells are cardiomyocytes.Results Beating myocardial cells were clearly observed after 48 hours of culture and 96.7% of cultured cells were live myocardial cells.Conclusion The protocol for isolation and cultivation of myocardial cells from neonatal mice are optimized.

关 键 词：乳小鼠 心肌细胞 原代培养 

分 类 号：Q2-33[生物学—细胞生物学]