Robust, Long-Term Culture of Endoderm-Derived Hepatic Organoids for Disease Modeling
Robust, Long-Term Culture of Endoderm-Derived Hepatic Organoids for Disease Modeling
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DOI：
10.1016/j.stemcr.2019.08.007
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发表时间：
2019-10-08
影响因子：
5.9
通讯作者：
Erdal, Esra
中科院分区：
文献类型：
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作者：
Akbari, Soheil;Sevinc, Gulben Gurhan;Erdal, Esra
类器官技术已成为模拟肝脏疾病、进行药物筛选以及个性化治疗的一种强大的新兴工具。然而，这些应用在以可重复且高效的方式产生功能性肝细胞的能力方面存在局限性。在此，我们利用人诱导多能干细胞（iPSC）衍生的上皮细胞黏附分子（EpCAM）阳性内胚层细胞作为中间产物，生成并表征了肝类器官（eHEPO）培养系统。eHEPO可在2周内产生，并能长期扩增（>16个月），且向成熟肝细胞分化的能力没有任何损失。从患者特异性iPSC开始，我们对1型瓜氨酸血症进行了建模，这是一种由精氨琥珀酸合成酶（ASST）基因突变引起的尿素循环障碍。在eHEPO中，疾病相关的氨蓄积表型可通过野生型ASS1基因的过表达而逆转，这也表明该模型易于进行基因操作。因此，eHEPO是以快速且高效的方式产生功能性肝类器官的极佳的无限细胞来源。
Organoid technologies have become a powerful emerging tool to model liver diseases, for drug screening, and for personalized treatments. These applications are, however, limited in their capacity to generate functional hepatocytes in a reproducible and efficient manner. Here, we generated and characterized the hepatic organoid (eHEPO) culture system using human induced pluripotent stem cell (iPSC)-derived EpCAM-positive endodermal cells as an intermediate. eHEPOs can be produced within 2 weeks and expanded long term (>16 months) without any loss of differentiation capacity to mature hepatocytes. Starting from patient-specific iPSCs, we modeled citrullinemia type 1, a urea cycle disorder caused by mutations in the argininosuccinate synthetase (ASST) enzyme. The disease-related ammonia accumulation phenotype in eHEPOs could be reversed by the overexpression of the wild-type ASS1 gene, which also indicated that this model is amenable to genetic manipulation. Thus, eHEPOs are excellent unlimited cell sources to generate functional hepatic organoids in a fast and efficient manner.