Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku, Fukuura 3-9, Yokohama, Kanagawa 236-0004, Japan; Advanced Medical Research Center, Yokohama City University, Kanazawa-ku, Fukuura 3-9, Yokohama, Kanagawa 236-0004, Japan; PRESTO, Japan Science and Technology Agency, 4-1-8, Honcho, Kawaguchi, Saitama 332-0012, Japan; Division of Gastroenterology, Hepatology, and Nutrition and Developmental Biology, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA; Center for Stem Cell and Organoid Medicine (CuSTOM), Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku, Fukuura 3-9, Yokohama, Kanagawa 236-0004, Japan.
Cell Rep. 2017 Dec 5;21(10):2661-2670. doi: 10.1016/j.celrep.2017.11.005.
Organoid technology provides a revolutionary paradigm toward therapy but has yet to be applied in humans, mainly because of reproducibility and scalability challenges. Here, we overcome these limitations by evolving a scalable organ bud production platform entirely from human induced pluripotent stem cells (iPSC). By conducting massive "reverse" screen experiments, we identified three progenitor populations that can effectively generate liver buds in a highly reproducible manner: hepatic endoderm, endothelium, and septum mesenchyme. Furthermore, we achieved human scalability by developing an omni-well-array culture platform for mass producing homogeneous and miniaturized liver buds on a clinically relevant large scale (>10). Vascularized and functional liver tissues generated entirely from iPSCs significantly improved subsequent hepatic functionalization potentiated by stage-matched developmental progenitor interactions, enabling functional rescue against acute liver failure via transplantation. Overall, our study provides a stringent manufacturing platform for multicellular organoid supply, thus facilitating clinical and pharmaceutical applications especially for the treatment of liver diseases through multi-industrial collaborations.
类器官技术为治疗提供了革命性的范例，但尚未在人类中应用，主要是因为存在可重复性和可扩展性挑战。在这里，我们通过从人类诱导多能干细胞 (iPSC) 中完全进化出可扩展的类器官芽生产平台来克服这些限制。通过进行大规模的“反向”筛选实验，我们确定了三个祖细胞群体，它们可以以高度可重复的方式有效地产生肝芽：肝内胚层、内皮细胞和隔膜间充质。此外，我们通过开发一种全孔阵列培养平台来实现人类的可扩展性，该平台可大规模生产均质和微型化的肝芽，规模达到临床相关的较大规模 (>10)。完全由 iPSC 产生的血管化和功能性肝组织通过匹配阶段的发育祖细胞相互作用显著改善了随后的肝功能化，从而通过移植实现对急性肝衰竭的功能挽救。总体而言，我们的研究为多细胞类器官供应提供了严格的制造平台，从而通过多产业合作促进了临床和药物应用，特别是用于治疗肝脏疾病。
