Tsoi Chantelle, Deng Ruixia, Kwok Maxwell, Yan Bin, Lee Carrie, Li Hung Sing, Ma Chloe Ho Yi, Luo Ruibang, Leung Kam Tong, Chan Godfrey Chi-Fung, Chow Larry Ming-Cheung, Poon Ellen N
Centre for Cardiovascular Genomics and Medicine, Lui Che Woo Institute of Innovative Medicine, The Chinese University of Hong Kong (CUHK), Shatin, Hong Kong SAR, China.
Hong Kong Hub of Paediatric Excellence (HK HOPE), CUHK, Shatin, Hong Kong SAR, China.
Front Mol Biosci. 2022 Mar 24;9:714008. doi: 10.3389/fmolb.2022.714008. eCollection 2022.
Inefficient differentiation and insufficient maturation are barriers to the application of human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) for research and therapy. Great strides have been made to the former, and multiple groups have reported cardiac differentiation protocol that can generate hPSC-CMs at high efficiency. Although many such protocols are based on the modulation of the WNT signaling pathway, they differ in their timing and in the WNT inhibitors used. Little is currently known about whether and how conditions of differentiation affect cardiac maturation. Here we adapted multiple cardiac differentiation protocols to improve cost-effectiveness and consistency, and compared the properties of the hPSC-CMs generated. Our results showed that the schedule of differentiation, but not the choice of WNT inhibitors, was a critical determinant not only of differentiation efficiency, which was expected, but also CM maturation. Among cultures with comparable purity, hPSC-CMs generated with different differentiation schedules vary in the expression of genes which are important for developmental maturation, and in their structural, metabolic, calcium transient and proliferative properties. In summary, we demonstrated that simple changes in the schedule of cardiac differentiation could promote maturation. To this end, we have optimized a cardiac differentiation protocol that can simultaneously achieve high differentiation efficiency and enhanced developmental maturation. Our findings would advance the production of hPSC-CMs for research and therapy.
低效分化和成熟不足是人类多能干细胞（hPSC）来源的心肌细胞（CMs）用于研究和治疗的应用障碍。在前者方面已经取得了很大进展，多个研究小组报告了能够高效产生hPSC-CMs的心脏分化方案。尽管许多此类方案基于WNT信号通路的调控，但它们在时间安排和所使用的WNT抑制剂方面存在差异。目前对于分化条件是否以及如何影响心脏成熟知之甚少。在这里，我们调整了多种心脏分化方案以提高成本效益和一致性，并比较了所产生的hPSC-CMs的特性。我们的结果表明，分化时间表而非WNT抑制剂的选择不仅是分化效率（这是预期的）的关键决定因素，也是CM成熟的关键决定因素。在纯度相当的培养物中，不同分化时间表产生的hPSC-CMs在对发育成熟重要的基因表达以及其结构、代谢、钙瞬变和增殖特性方面存在差异。总之，我们证明了心脏分化时间表的简单改变可以促进成熟。为此，我们优化了一种心脏分化方案，该方案可以同时实现高分化效率和增强的发育成熟。我们的发现将推动用于研究和治疗的hPSC-CMs的生产。
