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One-step 3D Differentiation of Cardiac Cells from Adult Human iPS Cells


詳細技術說明

Current state-of-the-art human iPS cell differentiation into contracting cardiomyocytes (CMs)follows either a 2D sheet approach or employs 3D cell aggregates (embryoid bodies). Both ofthese techniques require cells to be dissociated for downstream applications, including formationof engineered heart tissues and potential clinical applications, resulting in multiple cell handlingsteps and high levels of cell death. This method cultures and differentiates human iPS cells intocardiomyocytes directly in PEG-fibrinogen, a biodegradable hybrid biomaterial. 2D culturing ordifferentiating prior to 3D hydrogel formation is not required. Cardiomyocytes generated by thismethod retain their contractile function at ~140 beats/minute for several months and haveobservable aligned sarcomeres, Z-bands, H-bands, and t-tubules. Sarcomeric α-actinin, Cx43,and Nkx2.5 have been detected throughout the entire tissue (Panels c & d below). This methodcan be used for more rapid, cost effective production of 3D cardiomyocyte structures for drugscreening, engineered cardiac tissue formation, or other uses.


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