摘要
目的:探讨新生大鼠心肌细胞的分离和培养方法。方法:应用0.0625%的胰蛋白酶重复消化出生第2 d乳鼠的心肌组织多次,收集的细胞用含10%胎牛血清的DMEM培养基中和,用差速贴壁分离法分离,在以溴脱氧尿嘧啶(Brdu)纯化心肌细胞后置CO2培养箱孵育7 d。结果:分离1只乳鼠获得的心肌细胞产量约为140万个,且有活力心肌细胞占90%以上;培养4～6h的乳鼠心肌细胞开始贴壁生长,12～24h明显增殖,3～4 d后细胞融合成片;心肌细胞由圆形变为梭形、星形、多角形,并出现自发性节律性搏动。结论:本研究应用的心肌细胞原代培养方法可获得高产量、高活力的心肌细胞,是一种可靠的心肌细胞培养方法。
Objective： To explore the primary cultures method of neonatal rat cardiomyoeytes. Methods： Cardiac muscle of two-day neonate rattus was digested repeatedly by 0. 0625 % trypsin. The cardiocytes Were collected in DMEM medium, which contains 10% fetal bovine serum. Myocardial cells were isolated with the technique of differential anchoring velocity with 90 min and were pured by Brdu, The cardiocytes were placed into CO2 incubator to incubate seven days after their purification. Results： A total of 1.4 million cardiocytes can be obtained from one rat and the vital cell was over 90%; Cardiocytes began to adhere and grow after 4- 6 h, to obviously proliferation after 12-24 h, and to converge together after3-4 days. The cardiocytes showed spherical, fusiform and polygon shape in the visual field of inverted microscope. All cells stretched out parapodium and beated spontaneously and rhythmically. Conclusion： A great quantity and high survival rate cardiocytes can be effectively and quickly cultured by this method.
出处
《心血管康复医学杂志》
CAS
2009年第2期125-128,F0003,共5页
Chinese Journal of Cardiovascular Rehabilitation Medicine
关键词
心肌
细胞培养技术
大鼠
Myocardium
Cell culture techniques
Rat
