新生大鼠心房肌细胞的原代培养和快速电场起搏模型的建立  

Primary culture of atrial myocytes of neonatal rats and the construction of rapid electrical pacing model

作　　者：王伟[1] 程伟[1] 肖颖彬[1] 

机构地区：[1]第三军医大学附属新桥医院全军心血管外科中心,重庆市400037

出　　处：《实用医学杂志》2009年第21期3563-3565,共3页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:30600252)

摘　　要：目的:探讨心房肌细胞原代培养和快速电场起搏模型的建立方法,为对房颤早期电重构的研究奠定基础。方法:采用1周左右的大鼠,取左、右心房,胰蛋白酶结合Ⅱ型胶原酶消化细胞,利用2次差异性贴壁技术及加用Brdu纯化心房肌细胞,观察细胞形态以及结合免疫细胞化学检测心房肌细胞特异性表达的α-肌动蛋白鉴定心房肌细胞。结果:可得到高存活率和高纯度的心房肌细胞,经免疫细胞化学鉴定,90%以上培养细胞α-肌动蛋白抗体染色阳性。快速电场起搏24h后细胞存活率无显著降低。结论:分离培养大鼠心房肌细胞时,低浓度的胰酶和胶原酶合用,并用2次差速贴壁,可提高原代心肌培养的存活率和纯度,初步建立了快速起搏的房颤模型,为房颤的进一步研究奠定基础。Objective To establish the methods for primary culture of atrial myocytes and for construction of rapid electrical pacing model. Methods Specimens of atria were resected from one-week-old rats and digested by trypsogen and collagenase Ⅱ. Then the atrial myocytes were purified by technique of twice differential platting plus Brdu and identified by immunocytochemical analysis of α-aetin. Results Atrial myocytes with high survival rate and high purity were obtained successfully, and after being identified by immunocytochemical analysis, over 90% of these cells were stained positively for α-actin. There was no significant change in survival rate of atrial myoeytes after rapid electrical pacing. Conclusions Low density pancreatic enzyme and collagenase Ⅱ with the technique of differential anchoring velocity to isolate and culture atrial myocytes can improve the survival rate and purity of them. Cellular model of rapid electrical field pacing was established successfully.

关 键 词：肌细胞 心脏 原代培养 心房纤颤 快速起搏 大鼠 

分 类 号：R-332[医药卫生]