摘要
目的研究多次差速贴壁并联合应用5-溴脱氧尿苷（5-BrdU）对分离的心肌细胞纯度及活性的影响。方法以酶消化法分离乳鼠心肌细胞，分别进行1—3次差速贴壁并联用或不联用5-BrdU以纯化心肌细胞。正常培养3d后，观察细胞的形态特征。用流式细胞技术分析各实验组心肌细胞的纯度。以台盼蓝染色、MTT比色法及细胞线粒体内膜功能检测（流式细胞术）来反映各实验组心肌细胞的活性。结果随着差速贴壁次数的增加，心肌细胞得以纯化，但细胞的活性有所下降。联用5-BrdU后细胞纯度进一步提高，且对细胞的活性无明显影响；其中两次差速贴壁并联合应用5-BrdU后，可获得纯度高、活性好的心肌细胞。结论两次差速贴壁并联合应用5-BrdU为纯化心肌细胞的有效方法。
AIM To investigate the effects of repeatedly differential attachment technique combined with 5-BrdU on the purity and the activity of isolated cardiomyocytes. METHODS Sucking rat＇s cardiomyo-cytes were digested and isolated and then purified by differential attachment once to three times combined with or without 5-BrdU respectively. After the cardiomyocytes were cultured for 3 days, the morphological features of cultured cells were observed under inverted phase contrast microscope. The purity and the activity of eardiomyocytes were detected by flow eytometer （ FCM）, trypan-blue staining and MTT colorimetry. RESULTS The purity of cardiomyocytes was elevated with increased differential attachment times, but its activity decreased. After combined with 5-BrdU, the cellular purity and activity rose further and the effect of differential attachment twice combined with 5-BrdU was better. CONCLUSION The differential attachment twice combined with 5-BrdU is an effective method for the purification of isolated cardiomyocytes.
出处
《心脏杂志》
CAS
2008年第5期566-569,共4页
Chinese Heart Journal
基金
湖北省教育厅重点项目资助(D200624006)
