Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study
Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study
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DOI：
10.1186/s13287-020-01914-1
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发表时间：
2020-09-11
影响因子：
7.5
通讯作者：
Sprando, Robert L.
中科院分区：
文献类型：
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作者：
Gao, Xiugong;Li, Rong;Sprando, Robert L.
背景来源于人诱导多能干细胞（iPSC）的肝细胞样细胞（HLC）在毒理学和再生医学方面具有广阔的应用前景。以前的肝细胞分化的努力主要依赖于使用生长因子（GF）在体外条件下重演发育信号。最近，使用小分子（SM）已成为一个有吸引力的工具，以诱导细胞命运的转变，由于其在质量和成本方面的优势。然而，使用SM衍生的HLCs还没有得到很好的表征，特别是在transcriptome level.MethodsHLCs从人iPSCs使用的协议，只涉及SM和其特征在于使用全基因组microarrays. ResultsHLCs衍生使用SM协议（HLCs_SM）显示特定的肝标志物的表达，并表现出关键的肝功能。SM驱动的分化的转录组学分析定义了肝细胞分化轨迹，并表征了肝细胞分化的主要阶段中一些关键标志基因的表达。此外，使用CellNet（一种生物信息学工具，量化工程化细胞群与其靶细胞类型的相似程度）对HLC_SM进行评分，并与原代人肝细胞（PHH）、成人肝脏组织、胎儿肝脏组织、使用GF分化的HLC（HLC_GF）进行比较。），和市售的HLC。与HLC_GF相似，HLC_SM显示胎儿和成人肝细胞的混合表型，并且与PHH相比具有相对低的代谢酶、转运蛋白和核受体表达。最后，在HLC_SM相比，HLC_GF和PHH差异表达的基因进行了分析，以确定可能被操纵，以提高HLCs的分化的途径和上游转录调节因子。并提出了进一步改善肝细胞分化的方法，以获得可用于毒理学研究的更成熟的HLC。
BackgroundHepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) hold great promise in toxicological applications as well as in regenerative medicine. Previous efforts on hepatocyte differentiation have mostly relied on the use of growth factors (GFs) to recapitulate developmental signals under in vitro conditions. Recently, the use of small molecules (SMs) has emerged as an attractive tool to induce cell fate transition due to its superiority in terms of both quality and cost. However, HLCs derived using SMs have not been well characterized, especially on the transcriptome level.MethodsHLCs were differentiated from human iPSCs using a protocol that only involves SMs and characterized by transcriptomic analysis using whole genome microarrays.ResultsHLCs derived using the SM protocol (HLC_SM) displayed specific hepatic marker expression and demonstrated key hepatic functions. Transcriptomic analysis of the SM-driven differentiation defined a hepatocyte differentiation track and characterized the expression of some key marker genes in major stages of hepatocyte differentiation. In addition, HLC_SM were scored with CellNet, a bioinformatics tool quantifying how closely engineered cell populations resemble their target cell type, and compared to primary human hepatocytes (PHHs), adult liver tissue, fetal liver tissue, HLCs differentiated using GFs (HLC_GF), and commercially available HLCs. Similar to HLC_GF, HLC_SM displayed a mixed phenotype of fetal and adult hepatocytes and had relatively low expression of metabolic enzymes, transporters, and nuclear receptors compared to PHHs. Finally, the differentially expressed genes in HLC_SM compared to HLC_GF and to PHHs were analyzed to identify pathways and upstream transcription regulators which could potentially be manipulated to improve the differentiation of HLCs.ConclusionsOverall, the present study demonstrated the usefulness of the SM-based hepatocyte differentiation method, offered new insights into the molecular basis of hepatogenesis and associated gene regulation, and suggested ways for further improvements in hepatocyte differentiation in order to obtain more mature HLCs that could be used in toxicological studies.