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Zhang, Miao; Schulte, Jan Sebastian; Heinick, Alexander; Piccini, Ilaria; Rao, Jyoti; Quaranta, Roberto; Zeuschner, Dagmar; Malan, Daniela; Kim, Kee‐Pyo; Röpke, Albrecht; Sasse, Philipp; Araúzo‐Bravo, Marcos; Seebohm, Guiscard; Schöler, Hans; Fabritz, Larissa; Kirchhof, Paulus; Müller, Frank Ulrich; Greber, Boris
Stem cells (Dayton, Ohio), 20/May , Volume: 33, Issue: 5Journal Article
Directed cardiac differentiation of human pluripotent stem cells (hPSCs) enables disease modeling, investigation of human cardiogenesis, as well as large‐scale production of cardiomyocytes (CMs) for translational purposes. Multiple CM differentiation protocols have been developed to individually address specific requirements of these diverse applications, such as enhanced purity at a small scale or mass production at a larger scale. However, there is no universal high‐efficiency procedure for generating CMs both in two‐dimensional (2D) and three‐dimensional (3D) culture formats, and undefined or complex media additives compromise functional analysis or cost‐efficient upscaling. Using systematic combinatorial optimization, we have narrowed down the key requirements for efficient cardiac induction of hPSCs. This implied differentiation in simple serum and serum albumin‐free basal media, mediated by a minimal set of signaling pathway manipulations at moderate factor concentrations. The method was applicable both to 2D and 3D culture formats as well as to independent hPSC lines. Global time‐course gene expression analyses over extended time periods and in comparison with human heart tissue were used to monitor culture‐induced maturation of the resulting CMs. This suggested that hPSC‐CMs obtained with our procedure reach a rather stable transcriptomic state after approximately 4 weeks of culture. The underlying gene expression changes correlated well with a decline of immature characteristics as well as with a gain of structural and physiological maturation features within this time frame. These data link gene expression patterns of hPSC‐CMs to functional readouts and thus define the cornerstones of culture‐induced maturation. Stem Cells 2015;33:1456–1469
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