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.
Methods Mol Biol. 2022;2454:145-161. doi: 10.1007/7651_2021_395.
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.
人类诱导多能干细胞（hiPSCs）可在体外无限规模扩增，并分化为各种器官样细胞，包括心肌细胞（CMs）。先进的方案通过控制生长因子的应用来塑造多能干细胞的分化过程。调控 Wnt 信号通路可以有效地将 hiPSCs 诱导为心肌细胞（hiPSC-CMs），并使用小分子化合物替代天然生长因子。在这里，我们描述了一种可扩展生成 hiPSC-CMs 的改良方案，该方案操纵 Wnt 信号的豪猪和 Tankyrase 亚途径，以紧密抑制非经典 Wnt 信号。该方法可在搅拌生物反应器培养物中实现 hiPSC-CMs 的分化效率达到 87±0.9%，每 100 mL 无血清心脏分化培养基可产生约 7000 万 hiPSC-CMs。该分化方案易于从 3D 培养适应到 2D 培养，反之亦然，并且已被证明可与不同的 hiPSC 系配合使用。
