1] Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California, USA. [2] Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA. [3] Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California, USA.
Department of Applied Physics, Stanford University School of Medicine, Stanford, California, USA.
Nat Methods. 2014 Aug;11(8):855-60. doi: 10.1038/nmeth.2999. Epub 2014 Jun 15.
Existing methods for human induced pluripotent stem cell (hiPSC) cardiac differentiation are efficient but require complex, undefined medium constituents that hinder further elucidation of the molecular mechanisms of cardiomyogenesis. Using hiPSCs derived under chemically defined conditions on synthetic matrices, we systematically developed an optimized cardiac differentiation strategy, using a chemically defined medium consisting of just three components: the basal medium RPMI 1640, L-ascorbic acid 2-phosphate and rice-derived recombinant human albumin. Along with small molecule-based induction of differentiation, this protocol produced contractile sheets of up to 95% TNNT2(+) cardiomyocytes at a yield of up to 100 cardiomyocytes for every input pluripotent cell and was effective in 11 hiPSC lines tested. This chemically defined platform for cardiac specification of hiPSCs will allow the elucidation of cardiomyocyte macromolecular and metabolic requirements and will provide a minimal system for the study of maturation and subtype specification.
现有的人类诱导多能干细胞（hiPSC）心脏分化方法虽然高效，但需要复杂、未定义的培养基成分，这阻碍了对心肌发生分子机制的进一步阐明。使用在合成基质上通过化学定义条件获得的 hiPSC，我们系统地开发了一种优化的心脏分化策略，使用仅由三种成分组成的化学定义培养基：基础培养基 RPMI 1640、L-抗坏血酸 2-磷酸和源自大米的重组人白蛋白。通过基于小分子的诱导分化，该方案产生了高达 95% TNNT2(+)心肌细胞的收缩片，每输入的多能细胞的产量高达 100 个心肌细胞，在测试的 11 个 hiPSC 系中均有效。这种用于 hiPSC 心脏特化的化学定义平台将允许阐明心肌细胞的大分子和代谢需求，并为研究成熟和亚型特化提供一个最小系统。
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