Highly efficient and rapid generation of human pluripotent stem cells by chemical reprogramming (WGBS). Highly efficient and rapid generation of human pluripotent stem cells by chemical reprogramming (WGBS)

We recently demonstrated the chemical reprogramming of human somatic cells to pluripotent stem cells, which provides a fundamentally new approach for cell fate manipulation. However, the utility of this chemical approach is currently hampered by the slow kinetics. Here, by screening for new small-molecule boosters and systematically optimizing the original chemical reprogramming condition, we have established a robust, chemically defined protocol. With this new protocol, the entire chemical reprogramming process is remarkably accelerated, reducing the time needed from ~50 days to a minimum of 16 days. Moreover, this new protocol enables highly reproducible and efficient generation of human pluripotent stem cells from all 17 tested donors with a reprogramming efficiency of up to 31%. Our method provides a unique and powerful strategy for human cell fate manipulation and pluripotent stem cell manufacturing for cell-based therapeutic applications. Overall design: To characterize human chemically induced pluripotent stem cells (hCiPSCs), hCiPSCs were analyzed and compared to human somatic cells and human embryonic stem cells (hES) by WGBS.

本团队近期成功实现人类体细胞向多能干细胞的化学重编程，为细胞命运调控提供了全新的策略。然而该化学方法的应用目前受限于其缓慢的反应动力学过程。本研究通过筛选新型小分子增效剂，并系统性优化原始化学重编程培养条件，建立了一套稳健且化学成分明确的重编程方案。采用该新型方案，整个化学重编程流程得到显著加速，将所需时间从约50天缩短至最短16天。此外，该方案可实现高效且高重现性的多能干细胞制备：针对全部17个受试细胞供体，均可成功诱导获得人类多能干细胞，重编程效率最高可达31%。本方法为人类细胞命运调控以及面向细胞治疗应用的多能干细胞规模化制备提供了独特且高效的策略。
实验整体设计：为表征人类化学诱导多能干细胞 (human chemically induced pluripotent stem cells, hCiPSCs)，本研究通过全基因组亚硫酸氢盐测序 (Whole Genome Bisulfite Sequencing, WGBS) 对hCiPSCs、人类体细胞及人类胚胎干细胞 (human embryonic stem cells, hES) 进行了分析与比较。