Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.
Liver Cell Biology research group, Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel (VUB), Brussels, Belgium.
Nat Protoc. 2021 May;16(5):2542-2563. doi: 10.1038/s41596-021-00509-1. Epub 2021 Apr 16.
Hepatic stellate cells (HSCs) are nonparenchymal liver cells responsible for extracellular matrix homeostasis and are the main cells involved in the development of liver fibrosis following injury. The lack of reliable sources of HSCs has hence limited the development of complex in vitro systems to model liver diseases and toxicity. Here we describe a protocol to differentiate human induced pluripotent stem cells (iPSCs) into hepatic stellate cells (iPSC-HSCs). The protocol is based on the addition of several growth factors important for liver development sequentially over 12 d. iPSC-HSCs present phenotypic and functional characteristics of primary HSCs and can be expanded or frozen and used to perform high-throughput in vitro studies. We also describe how to coculture iPSC-HSCs with hepatocytes, which self-assemble into three-dimensional (3D) hepatic spheroids. This protocol enables the generation of HSC-like cells for in vitro modeling and drug screening studies.
肝星状细胞（HSCs）是负责细胞外基质稳态的非实质细胞，是损伤后肝纤维化发展的主要细胞。缺乏可靠的 HSCs 来源，因此限制了用于模拟肝脏疾病和毒性的复杂体外系统的发展。本文描述了一种将人诱导多能干细胞（iPSCs）分化为肝星状细胞（iPSC-HSCs）的方案。该方案基于在 12 天内顺序添加几种对肝脏发育很重要的生长因子。iPSC-HSCs 表现出原代 HSCs 的表型和功能特征，可进行扩增或冷冻，并用于进行高通量体外研究。我们还描述了如何将 iPSC-HSCs 与肝细胞共培养，肝细胞会自我组装成三维（3D）肝球体。该方案可用于生成用于体外建模和药物筛选研究的 HSC 样细胞。
