Generation of hepatobiliary organoids from human induced pluripotent stem cells
Generation of hepatobiliary organoids from human induced pluripotent stem cells
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从人类诱导多能干细胞生成肝胆类器官
DOI：
10.1016/j.jhep.2018.12.028
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发表时间：
2019-06-01
影响因子：
25.7
通讯作者：
Xu, Anlong
中科院分区：
文献类型：
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作者：
Wu, Fenfang;Wu, Di;Xu, Anlong
背景与目的：人类诱导多能干细胞（hiPSC）衍生的肝脏模型系统有潜力克服临床应用中供体短缺的问题，并成为药物开发的模型。虽然有几种策略可用于生成肝脏微组织，但很少有策略成功地从hipsc中生成具有肝胆结构的肝类器官。方法：在分化阶段1和II（第1-15天），在肝分化培养基中加入25%的mTeSRTM培养基，诱导内胚层和中胚层分化，随后肝和胆道共分化。在第三阶段（第15-45天），成熟培养基中加入10%胆固醇+ MIX，促进肝胆类器官的形成和成熟。类器官的表型和功能通过体外和体内的特异性标记和多种功能测定来确定。结果：在该系统中，hiPSCs被诱导形成3D肝胆类器官，并在一定程度上以平行方式再现了早期肝发生的关键方面。类器官显示出一系列的功能属性。具体而言，诱导的肝细胞样细胞可以摄取吲哚菁绿，积累脂质和糖原，并表现出适当的分泌能力（白蛋白和尿素）和药物代谢能力（CYP3A4活性和诱导性）；系统中的胆道结构显示γ谷氨酰转移酶活性和排出罗丹明和储存胆汁酸的能力。此外，移植到免疫缺陷小鼠体内后，类器官存活超过8周。结论：这是首次从hipsc中生成功能性肝胆类器官。类器官模型将有助于肝脏发育的分子机制的体外研究，并在肝脏疾病的治疗中具有重要的潜力。摘要：在此，我们建立了一个体外生成人诱导多能干细胞衍生的功能性肝胆类器官的系统，不需要任何外源细胞或基因操作。在某种程度上，该模型能够以平行的方式概括肝胆器官发生的几个关键方面，对药物开发和肝移植具有很大的希望。(C) 2019欧洲肝脏研究协会。Elsevier B.V.出版
Background & Aims: Human induced pluripotent stem cell (hiPSC)-derived liver modeling systems have the potential to overcome the shortage of donors for clinical application and become a model for drug development. Although several strategies are available to generate hepatic micro-tissues, few have succeeded in generating a liver organoid with hepatobiliary structure from hiPSCs.Methods: At differentiation stages I and II (day 1-15), 25% of mTeSRTM culture medium was added to hepatic differentiation medium to induce endodermal and mesodermal commitment and thereafter hepatic and biliary co-differentiation. At stage III (day 15-45), 10% cholesterol+ MIX was added to the maturation medium to promote the formation and maturation of the hepatobiliary organoids. Phenotypes and functions of organoids were determined by specific markers and multiple functional assays both in vitro and in vivo.Results: In this system, hiPSCs were induced to form 3D hepatobiliary organoids and to some extent recapitulated key aspects of early hepatogenesis in a parallel fashion. The organoids displayed a series of functional attributes. Specifically, the induced hepatocyte-like cells could take up indocyanine green, accumulate lipid and glycogen, and displayed appropriate secretion ability (albumin and urea) and drug metabolic ability (CYP3A4 activity and inducibility); the biliary structures in the system showed gamma glutamyltransferase activity and the ability to efflux rhodamine and store bile acids. Furthermore, after transplantation into the immune-deficient mice, the organoids survived for more than 8 weeks.Conclusion: This is the first time that functional hepatobiliary organoids have been generated from hiPSCs. The organoid model will be useful for in vitro studies of the molecular mechanisms of liver development and has important potential in the therapy of liver diseases.Lay summary: Herein, we established a system to generate human induced pluripotent stem cell-derived functional hepatobiliary organoids in vitro, without any exogenous cells or genetic manipulation. To some extent this model was able to recapitulate several key aspects of hepatobiliary organogenesis in a parallel fashion, holding great promise for drug development and liver transplantation. (C) 2019 European Association for the Study of the Liver. Published by Elsevier B.V.