The research on human hepatobiliary development and disorders has been constrained by minimal access to human fetal tissue, and low accuracy of animal models. To overcome this problem, we have established a system for the differentiation of human pluripotent stem cells (hPSCs) into functional hepatobiliary organoids (HBOs). We have previously reported that our 45-d approach closely mimics key stages of hepatobiliary development, starting with the differentiation of hiPSC into endoderm and a small part of mesoderm, and subsequently into hepatoblast-like cells, followed by the parallel generation of hepatocyte-like cells and cholangiocyte-like cells, formation of immature HBO expressing early hepatic and biliary markers, and mature HBO displaying hepatobiliary functionality. In this study, we present an updated version of our previous protocol, which only needs 35 days to achieve maturation in vitro. Furthermore, a hepatobiliary culture medium is developed to functionally maintain the HBOs for more than 1.5 months. The capacity of this approach for producing large amounts of functional HBOs and enabling long-term culture in vitro holds promise for applications on developmental research, disease modeling, as well as screening of therapeutic agents.

中文翻译：

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从人多能干细胞生产功能性肝胆类器官。

人类肝胆发育和疾病的研究受到人类胎儿组织的最少接触和动物模型准确性低的限制。为了克服这个问题，我们建立了一个将人多能干细胞（hPSC）分化为功能性肝胆类器官（HBO）的系统。我们以前曾报道过，我们的45天方法紧密模拟了肝胆发育的关键阶段，首先是将hiPSC分化为内胚层和一小部分中胚层，然后分化为成肝细胞样细胞，然后平行生成肝细胞样细胞。细胞和胆管样细胞，形成早期肝和胆汁标志物的未成熟HBO，以及显示肝胆功能的成熟HBO。在这项研究中，我们介绍了先前协议的更新版本，体外。此外，开发了肝胆培养基以在功能上维持HBO超过1.5个月。这种方法产生大量功能性HBO的能力以及能够在体外长期培养的能力，有望在发展研究，疾病建模以及治疗药物筛选中得到应用。