论文部分内容阅读
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 investigate the method of isolation and culture of cardiomyocytes in vitro in mice and to establish an optimized method of culturing cardiomyocytes in vitro in mice. Methods The trypsin-Ⅱcollagenase mixture was used to digest the myocardium of the mice and differentiated into cardiomyocytes by differential adherent method. The characteristics of a-actin expressed specifically in cardiomyocytes were analyzed by immunohistochemical method Identify cardiomyocytes. Results After 48h, the cardiomyocytes adhered to the wall completely, and most of the adherent cells appeared pulsation, but no pulsing was observed. The purity of cardiomyocytes was 96.7% after a-actin cell immunohistochemistry. Conclusion This study optimizes the primary culture and isolation of cardiomyocytes from neonatal mice and lays the foundation for subsequent cell experiments.