Scalable Generation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

Autor:	Hamad S; Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Medical Faculty, Cologne, Germany.; Biology Department, Faculty of Science, Soran University, Soran, Kurdistan Region, Iraq., Derichsweiler D; Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Medical Faculty, Cologne, Germany., Hescheler J; Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Medical Faculty, Cologne, Germany., Pfannkuche K; Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Medical Faculty, Cologne, Germany. Kurt.Pfannkuche@uni-koeln.de.; Department of Pediatric Cardiology, University Clinics of Cologne, Cologne, Germany. Kurt.Pfannkuche@uni-koeln.de.; Center for Molecular Medicine (CMMC), University of Cologne, Cologne, Germany. Kurt.Pfannkuche@uni-koeln.de.; Marga-and-Walter-Boll Laboratory for Cardiac Tissue Engineering, University of Cologne, Cologne, Germany. Kurt.Pfannkuche@uni-koeln.de.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2454, pp. 145-161.
DOI:	10.1007/7651_2021_395
Abstrakt:	Human induced pluripotent stem cells (hiPSCs) can be expanded at limitless scale in vitro and give rise to various organotypic cells, cardiomyocytes (CMs) among them. Advanced protocols shape the differentiation process of pluripotent stem cells by controlled growth factor application. Modulating the Wnt signaling pathway is effective to direct hiPSCs to CMs (hiPSC-CMs) and native growth factors were replaced by small chemical compounds. Here, we describe a refined protocol for scalable generation of hiPSC-CMs that manipulates porcupine and tankyrase sub-pathways of Wnt signaling for tight inhibition of non-canonical Wnt signaling. The approach results in a differentiation efficiency toward hiPSC-CMs of 87 ± 0.9% in stirred bioreactor cultures and yields about 70 million hiPSC-CMs per 100 mL serum free cardiac differentiation medium. The differentiation protocol is easily adapted from 3D to 2D culture and vice versa and has been demonstrated to work with different hiPSC lines. (© 2021. Springer Science+Business Media, LLC.)
Databáze:	MEDLINE
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