Sohnis Family Research Laboratory for Cardiac Electrophysiology and Regenerative Medicine (Bruce Rappaport Faculty of Medicine), Technion-Israel Institute of Technology, Haifa, Israel.
Tissue Eng Part A. 2011 Apr;17(7-8):1027-37. doi: 10.1089/ten.TEA.2010.0235. Epub 2011 Jan 10.
Cardiomyocytes derived from induced pluripotent stem (iPS) cells hold great promise for basic and translational cardiovascular research. For the successful implementation of this unique technology, however, it is essential to establish efficient, reproducible, and safe strategies to produce cardiomyocytes in a scalable manner. The aim of the current study was to establish scalable bioprocess that allows direct embryoid bodies formation for the differentiation of murine iPS cells (generated without the oncogene c-Myc) into cardiomyocytes. The cardiomyocytes' structural, molecular, and functional properties were then compared to ones derived by the well-established static culture system. Similar gene expression patterns were observed in both differentiation systems with the sequential expression of mesoderm markers, cardiac transcription factors, and cardiomyocyte structural genes. Cells in the contracting embryoid bodies were stained positively for cardiac troponin-I, sarcomeric α-actinin, cardiac troponin-T, and connexin-43. Electrophysiological measurements using multielectrode array recordings demonstrated that the bioreactor-derived cardiomyocytes were functionally similar to static derived cardiomyocytes and responded appropriately to different drugs, including adrenergic and muscarinic agonists (isoproterenol and carbamylcholine, respectively) and the gap junction uncoupler heptanol. Our study describes, for the first time, a strategy for scalable differentiation of c-Myc-free iPS cells into cardiomyocytes with the appropriate molecular, structural, and functional properties. The result of this study should have important implications for several cardiovascular research areas and specifically for the emerging field of regenerative medicine.
心肌细胞来源于诱导多能干细胞（iPS），在基础和转化心血管研究中具有广阔的应用前景。然而，为了成功实施这项独特的技术，建立高效、可重复和安全的策略来规模化生产心肌细胞是至关重要的。本研究旨在建立一种可规模化的生物工艺，允许直接形成类胚体，从而将鼠源 iPS 细胞（无癌基因 c-Myc 产生）分化为心肌细胞。然后将心肌细胞的结构、分子和功能特性与通过成熟的静态培养系统获得的心肌细胞进行比较。在这两种分化系统中观察到相似的基因表达模式，表现为中胚层标记物、心脏转录因子和心肌细胞结构基因的顺序表达。在收缩的类胚体中的细胞对心肌肌钙蛋白 I、肌节α-肌动蛋白、心肌肌钙蛋白 T 和连接蛋白 43 呈阳性染色。使用多电极阵列记录进行的电生理测量表明，生物反应器衍生的心肌细胞在功能上与静态衍生的心肌细胞相似，并且对不同的药物（包括肾上腺素能和毒蕈碱激动剂（异丙肾上腺素和卡巴胆碱）和间隙连接解偶联剂庚醇）有适当的反应。本研究首次描述了一种将无 c-Myc 的 iPS 细胞规模化分化为具有适当分子、结构和功能特性的心肌细胞的策略。这项研究的结果应该对几个心血管研究领域具有重要意义，特别是对再生医学这一新兴领域。
