基础医学与临床 ›› 2009, Vol. 29 ›› Issue (9): 916-919.

• 研究论文 • 上一篇    下一篇

小鼠胚胎干细胞来源的心肌细胞系的建立

秦洁 薛兆英 郭新 桂耀庭 余振东 蔡志明   

  1. 北京大学深圳医院 北京大学深圳医院
  • 收稿日期:2008-11-27 修回日期:2009-01-24 出版日期:2009-09-20 发布日期:2009-09-20
  • 通讯作者: 蔡志明

Establishment of cardiomyocyte from mouse embryonic stem cell-derived embryoid bodies

Jie QIN, Zhao-ying XUE, Xin GUO, Yao-ting GUI, Zhen-dong YU, Zhi-ming CAI   

  1. Peking University ShenZhen Hospital Peking University ShenZhen Hospital
  • Received:2008-11-27 Revised:2009-01-24 Online:2009-09-20 Published:2009-09-20
  • Contact: Zhi-ming CAI

摘要: 目的 建立模拟细胞微环境、促进细胞分化的三维培养体系,以体外诱导和培养小鼠胚胎干细胞 (mES-D3)来源的心肌细胞。方法 将mES细胞通过悬浮培养获得拟胚体(EBs)后转入Matrigel? Matrix 3D培养体系中培养。用形态学观察计数收缩集落的数量和频率;用免疫组织化学和RT-PCR检测心肌细胞分化相关基因的表达。结果 在Matrigel?中培养5d左右,可观察到自发节律收缩的EBs,频率约为40~50次/min;继续培养3d左右,频率约为80~90次/min;到45d左右大部分集落最终收缩。分化为心肌细胞所表达的特异性蛋白cTnT, Desmin, Actin表达呈阳性,并表达心肌特异性基因cTnT, NKX2E, GATA-4和MLC-2v。结论 mES细胞来源的EBs在Matrigel?培养体系中能够分化为心肌细胞,籍此我们建立了一株细胞系。

关键词: 小鼠胚胎干细胞(mES), 拟胚体 (EBs), 分化, 心肌细胞

Abstract: Objective To establishment a defined three-dimensional culture system of cardiomyocytes from mouse embryonic stem cell-derived embryoid bodies (EBs). Methods The EBs were derived from mES by suspending culture and were plated to Matrigel? Matrix three-dimensional system. The mouse fibroblast cells served as negative control to identify genes expression of contracting EBs. Results The first beating was irregular after 5 days of culture, but became consistent after 3 days with at an average frequency of 80-100 beats per minute. mES expressed cardiac specific protein of cTnT,Desmin and Actin. The expression of genes associated cardiomyocytes differentiation as cTnT, NKX2E, GATA-4 and MLC-2v were detected by RT-PCR. Conclusion By using the Matrigel? three-dimensional system, mES-derived EBs could spontaneously differentiate to cardiomyocytes effectively, and possess necessary biological characters of cardiomyocytes .

Key words: Mouse embryonic stem(mES) cell, Embryoid Bodies(EBs), Differentiation, Cardiomyocytes