High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry

Subarna Bhattacharya; Paul W. Burridge; Erin M. Kropp; Sandra L. Chuppa; Wai-Meng Kwok; Joseph C. Wu; Kenneth R. Boheler; Rebekah L. Gundry

doi:10.3791/52010

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High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry

人类多能干细胞的高效分化为心肌细胞，并鉴定流式细胞仪

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13:13 min

September 23, 2014

DOI: 10.3791/52010-v

Subarna Bhattacharya*¹, Paul W. Burridge*², Erin M. Kropp¹, Sandra L. Chuppa¹, Wai-Meng Kwok³, Joseph C. Wu², Kenneth R. Boheler^4,5, Rebekah L. Gundry^1,6

¹Department of Biochemistry,Medical College of Wisconsin, ²Stanford Cardiovascular Institute,Stanford University School of Medicine, ³Department of Anesthesiology,Medical College of Wisconsin, ⁴Stem Cell and Regenerative Medicine Consortium, LKS Faculty of Medicine,Hong Kong University, ⁵Division of Cardiology,Johns Hopkins University School of Medicine, ⁶Cardiovascular Research Center, Biotechnology and Bioengineering Center,Medical College of Wisconsin

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该文章描述的详细方法通过选择性调节Wnt通路，随后通过流式细胞仪的参考标记分析来评估群体的均匀性和同一性的有效分化人多能干细胞分化为心肌细胞。

该程序的总体目标是从多能干细胞中产生高质量的心肌细胞培养物，并使用流式细胞术适当表征最终产物。为了实现这一点，生成了未分化多能干细胞的高质量单层培养物。然后通过使用小分子调节 wind 途径来诱导心肌生成。

接下来，使用专门针对每种抗体优化的条件对心肌细胞进行固定、渗透和染色。最后，通过流式细胞术分析细胞。最终，可以获得结果，显示培养物对心脏标志物 TNNI 3 和 IRX 4 高度阳性，并显示出类似心室的动作电位。

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细胞生物学第91 人类诱导多能干细胞流式细胞仪定向分化心肌细胞 IRX4 TNNI3 TNNT2 MCL2v MLC2a