小鼠胚胎干细胞来源的心肌细胞系的建立

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

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

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

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/Abstract

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

秦洁薛兆英郭新桂耀庭余振东蔡志明. 小鼠胚胎干细胞来源的心肌细胞系的建立[J]. 基础医学与临床, 2009, 29(9): 916-919.

Jie QIN; Zhao-ying XUE; Xin GUO; Yao-ting GUI; Zhen-dong YU; Zhi-ming CAI. Establishment of cardiomyocyte from mouse embryonic stem cell-derived embryoid bodies[J]. Basic & Clinical Medicine, 2009, 29(9): 916-919.

[1]	周杨周苗. 外泌体在骨再生领域的应用进展[J]. 基础医学与临床, 2019, 39(8): 1196-1199.
[2]	李静杨岩磊赵春华. 人脂肪间充质干细胞向限定性内胚层细胞分化体系的优化[J]. 基础医学与临床, 2019, 39(7): 954-960.
[3]	张源洲席雨鑫温馨李鸿珠. 硫化氢对衰老缺血心肌细胞保护作用的研究进展[J]. 基础医学与临床, 2019, 39(2): 252-255.
[4]	刘志强李妍杨小娟李慧宁韩远豪王晓玲汪涛. 小鼠肌源性干细胞在3种不同培养基中的增殖特征[J]. , 2019, 39(12): 1669-1674.
[5]	陈晓依王浩宇吕洋王海萍孙微李柔. PFT-α与BMP-2联合诱导大鼠BMMSCs分化为心肌样细胞[J]. 基础医学与临床, 2018, 38(8): 1088-1093.
[6]	任春贞骆亚莉刘永琦李程豪张艳辉王风梅卢志伟李玲. IL-6和TNF-α模拟炎性微环境促进人BMSCs向肿瘤相关成纤维细胞分化[J]. 基础医学与临床, 2018, 38(7): 901-906.
[7]	吴庆程阔菊姚浩旗黄河杨天德. 芒果苷对原代大鼠心肌细胞缺血缺氧凋亡的影响[J]. 基础医学与临床, 2018, 38(2): 169-173.
[8]	角建林唐薇郑红赵宏斌. 彝药恒古骨伤愈合剂缓解去势树鼩骨质疏松症[J]. 基础医学与临床, 2018, 38(12): 1686-1690.
[9]	周芳亮何迎春闫庆梓张凯强廖端芳. 大鼠骨髓间充质干细胞向血管平滑肌细胞分化过程中小窝蛋白-1表达增强[J]. 基础医学与临床, 2018, 38(11): 1517-1521.
[10]	赖星宇朱静田杰易勤陈雪妮. Akt在Islet-1促C3H10T1/2向心肌细胞特异分化过程中的作用[J]. 基础医学与临床, 2018, 38(1): 13-19.
[11]	崔申申解学敏王南宇吕湘. IFN-γ促进红系终末分化[J]. 基础医学与临床, 2017, 37(7): 963-969.
[12]	吕凤香康权仇超董姿杏罗庆. DLX1联合BMP9促进人骨肉瘤细胞向成骨分化[J]. 基础医学与临床, 2017, 37(6): 768-774.
[13]	代光明任磊陈虹刘文陈宇何小强刘伟涂小林黄伟. 下调骨细胞TGF-β/Smad4信号可抑制小鼠BMSCs成骨及破骨分化[J]. 基础医学与临床, 2017, 37(6): 786-791.
[14]	方连英王彦徐畅杜利清王芹刘强. 电离辐射诱导造血干细胞损伤的分子机制研究进展[J]. 基础医学与临床, 2017, 37(2): 256-260.
[15]	崔桂丽王瑞玲马琴闫佳敏王亚静. 自噬在高糖高脂引起H9C2心肌细胞系凋亡中的影响[J]. 基础医学与临床, 2017, 37(12): 1699-1705.