江苏大学学报（医学版）2025，Vol.35Issue(2)：161-165,5.DOI:10.13312/j.issn.1671-7783.y240093
新生小鼠原代心肌细胞分离及培养方法的改进
Improvement of method for isolation and culture of primary myocardial cells in neonatal mouse
摘要
Abstract
Objective:To improve the isolation and culture methods of primary cardiomyocytes in neonatal mouse to stably get high activity and purity of primary mouse cardiomyocytes.Methods:Hearts from 1-3-day-old neonatal mouse were harvested,and a two-step digestion method combining trypsin and type Ⅱ collagenase was used.Differential adherence was then employed to purify the cardiomyocytes,which were cultured in vitro in modified Dulbecco's Modified Eagle Medium(DMEM)containing 10%horse serum.The morphological characteristics and beating patterns of the cardiomyocytes were observed under an inverted phase-contrast microscope.Cell viability was assessed by using trypan blue staining,and cardiomyocytes were identified by immunofluorescence staining for cardiac troponin T(cTnT).Results:After 24 hours of culture,the primary neonatal mouse cardiomyocytes adhered to the culture dish and exhibited spontaneous beating.By 96 hours,the cells aggregated into clusters and displayed synchronous beating.The cell survival rate was 90.63%,and the purity of cardiomyocytes reached 93.17%.Conclusion:The modified isolation and culture methods used in this study obtained the primary neonatal mouse cardiomyocytes with high activity and purity.关键词
新生小鼠/心肌细胞/两步酶消化法/原代培养Key words
neonatal mouse/cardiomyocyte/two-step enzymatic digestion method/primary culture分类
医药卫生引用本文复制引用
罗洁,曹梦菲,付可威,吕书梅,袁伟..新生小鼠原代心肌细胞分离及培养方法的改进[J].江苏大学学报（医学版）,2025,35(2):161-165,5.基金项目
江苏省重点研发计划项目(BE2021694) （BE2021694）
