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Title:: Effects of repeatedly differential attachment technique combined with 5BrdU on the purity and the activity of isolated cardiomyocytes

作者:: 郎明健¹; 2; 郭敏¹; 曾秋棠¹; 闵新文²; 杨汉东²; 1.华中科技大学同济医学院附属协和医院心内科，湖北武汉 430022； 2.郧阳医学院附属东风医院心内科，湖北十堰 442008

Author(s):: LANG Mingjian¹; 2; GUO Min¹; ZENG Qiutang¹; MIN Xinwen²; YANG Handong2²; 1.Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei, China; 2.Department of Cardiology, Dongfeng Hospital, Yunyang Medical College, Shiyan 442008, Hubei, China

关键词:: 心肌细胞; 纯化; 差速贴壁技术; 5-溴脱氧尿苷

Keywords:: cardiomyocyte; purification; differential attachment technique; 5-BrdU

分类号:: R446.11

DOI:: -

文献标识码:: A

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3 d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。

Abstract:: 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 cardiomyocytes 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 cardiomyocytes were detected by flow cytometer (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.

参考文献/References

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差速贴壁联合应用5-BrdU对心肌细胞纯度及活力的影响(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

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