Asumda Faizal Z, Alzoubi Shadia, Padarath Kiyasha, Jones Kimya, Kolhe Ravindra, Mondal Ashis Kumar, Alptekin Ahmet, Zhi Wenbo, Lee Tae Jin, Huebert Robert C, Staff Nathan P, Roberts Lewis R, Kirkeby Lindsey A
Medical College of Georgia-Augusta University Department of Pediatrics, Augusta, GA, United States.
Medical College of Georgia-Augusta University Department of Pathology, Augusta, GA, United States.
Front Cell Dev Biol. 2025 Jun 26;13:1594340. doi: 10.3389/fcell.2025.1594340. eCollection 2025.
The growth factor and small molecule protocol are the two primary approaches for generating human induced pluripotent stem cell-derived hepatocyte-like cells (iPSC-HLCs). We compared the efficacy of the growth factor and small molecule protocols across fifteen different human iPSC lines. Morphological assessment, relative quantification of gene expression, protein expression and proteomic studies were carried out. HLCs derived from the growth factor protocol displayed mature hepatocyte morphological features including a raised, polygonal shape with well-defined refractile borders, granular cytoplasm with lipid droplets and/or vacuoles with multiple spherical nuclei or a large centrally located nucleus; significantly elevated hepatocyte gene and protein expression including AFP, HNF4A, ALBUMIN, and proteomic and metabolic features that are more aligned with a mature phenotype. HLCs derived from the small molecule protocol showed a dedifferentiated, proliferative phenotype that is more akin to liver tumor-derived cell lines. These experimental results suggest that HLCs derived from growth factors are better suited for studies of metabolism, biotransformation, and viral infection.
生长因子和小分子方案是生成人诱导多能干细胞来源的肝样细胞（iPSC-HLCs）的两种主要方法。我们比较了生长因子和小分子方案在15种不同人类iPSC系中的效果。进行了形态学评估、基因表达的相对定量、蛋白质表达和蛋白质组学研究。源自生长因子方案的HLCs表现出成熟肝细胞的形态特征，包括凸起的多边形形状、边界清晰的折光边缘、含有脂滴和/或空泡的颗粒状细胞质、多个球形核或位于中央的大核；肝细胞基因和蛋白质表达显著升高，包括甲胎蛋白（AFP）、肝细胞核因子4A（HNF4A）、白蛋白，以及在蛋白质组学和代谢特征上更符合成熟表型。源自小分子方案的HLCs表现出一种去分化的增殖表型，更类似于肝肿瘤衍生的细胞系。这些实验结果表明，源自生长因子的HLCs更适合用于代谢、生物转化和病毒感染的研究。
