Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media.
Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media.
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DOI：
10.1038/s41598-020-73908-1
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发表时间：
2020-10-21
影响因子：
4.6
通讯作者：
Taniguchi H
中科院分区：
文献类型：
--
作者：
Sekine K;Ogawa S;Tsuzuki S;Kobayashi T;Ikeda K;Nakanishi N;Takeuchi K;Kanai E;Otake Y;Okamoto S;Kobayashi T;Takebe T;Taniguchi H
类器官技术的进步已经扩大了可以应用基于人类诱导多能干细胞（iPSC）的再生医学的目标疾病和病症的数量;然而，类器官的大规模生产和化学成分确定的无动物源（CD-AOF）培养基和补充剂的开发是阻碍这些方法的临床适用性的未解决的问题。CD-AOF培养基和补充剂通过降低批间差异和病毒或毒素污染风险，确保培养系统的质量和重现性。我们先前通过模拟肝细胞、内皮细胞（EC）和间充质细胞（MC）之间的器官发生相互作用从iPSC产生了肝脏类器官，即iPSC-肝芽（iPSC-LB），并且最近报道了完全源自iPSC的iPSC-LB的大规模生产（所有iPSC-LB），这应该有助于它们的大规模生产用于治疗肝功能衰竭。然而，在先前的研究中，我们使用来源于动物的培养基进行分化，除了维持未分化的iPSC。因此，我们开发了CD-AOF培养基以产生所有iPSC-LB。我们首先开发了用于肝细胞、EC和阶段匹配的MC的CD-AOF培养基，即，横隔间充质（STM），2D培养物中。我们接下来通过在超低附着微凹板中孵育单个细胞类型来产生所有iPSC-LB。在体外和体内，使用CD-AOF培养基产生的所有iPSC-LB的肝功能与使用常规培养基产生的所有iPSC-LB的肝功能相当。此外，我们发现这种CD-AOF培养基可以用于几种细胞培养环境。综上所述，这些结果证明了适用于所有iPSC-LB的CD-AOF培养基的成功开发。本研究中开发的方案将促进所有iPSC-LB在肝脏疾病治疗中的临床适用性。
Advances in organoid technology have broadened the number of target diseases and conditions in which human induced pluripotent stem cell (iPSC)-based regenerative medicine can be applied; however, mass production of organoids and the development of chemically defined, animal origin-free (CD-AOF) media and supplements are unresolved issues that hamper the clinical applicability of these approaches. CD-AOF media and supplements ensure the quality and reproducibility of culture systems by lowering lot-to-lot variations and the risk of contamination with viruses or toxins. We previously generated liver organoids from iPSCs, namely iPSC-liver buds (iPSC-LBs), by mimicking the organogenic interactions among hepatocytes, endothelial cells (ECs), and mesenchymal cells (MCs) and recently reported the mass production of iPSC-LBs derived entirely from iPSCs (all iPSC-LBs), which should facilitate their large-scale production for the treatment of liver failure. However, in previous studies we used media originating from animals for differentiation except for the maintenance of undifferentiated iPSCs. Therefore, we developed a CD-AOF medium to generate all iPSC-LBs. We first developed a CD-AOF medium for hepatocytes, ECs, and stage-matched MCs, i.e., septum transversum mesenchyme (STM), in 2D cultures. We next generated all iPSC-LBs by incubating individual cell types in ultra-low attachment micro-dimple plates. The hepatic functions of all iPSC-LBs generated using the CD-AOF medium were equivalent to those of all iPSC-LBs generated using the conventional medium both in vitro and in vivo. Furthermore, we found that this CD-AOF medium could be used in several cell culture settings. Taken together, these results demonstrate the successful development of a CD-AOF medium suitable for all iPSC-LBs. The protocol developed in this study will facilitate the clinical applicability of all iPSC-LBs in the treatment of liver diseases.
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影响因子：
64.8
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影响因子：
64.8
作者：
Takasato, Minoru;Er, Pei X.;Little, Melissa H.
通讯作者：
Little, Melissa H.
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影响因子：
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作者：
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通讯作者：
Yamanaka, Shinya
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Tanabe, Koji;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya