Yanagihara Kana, Liu Yujung, Kanie Kei, Takayama Kazuo, Kokunugi Minako, Hirata Mitsuhi, Fukuda Takayuki, Suga Mika, Nikawa Hiroki, Mizuguchi Hiroyuki, Kato Ryuji, Furue Miho K
1 Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation , Health and Nutrition, Osaka, Japan .
2 Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University , Nagoya, Japan .
Stem Cells Dev. 2016 Dec 15;25(24):1884-1897. doi: 10.1089/scd.2016.0099. Epub 2016 Nov 8.
Functional hepatocytes derived from human pluripotent stem cells (hPSCs) have potential as tools for predicting drug-induced hepatotoxicity in the early phases of drug development. However, the propensity of hPSC lines to differentiate into specific lineages is reported to differ. The ability to predict low propensity of hPSCs to differentiate into hepatocytes would facilitate the selection of useful hPSC clones and substantially accelerate development of hPSC-derived hepatocytes for pharmaceutical research. In this study, we compared the expression of genes associated with hepatic differentiation in five hPSC lines including human ES cell line, H9, which is known to differentiate into hepatocytes, and an hPSC line reported with a poor propensity for hepatic differentiation. Genes distinguishing between undifferentiated hPSCs, hPSC-derived hepatoblast-like differentiated cells, and primary human hepatocytes were drawn by two-way cluster analysis. The order of expression levels of genes in undifferentiated hPSCs was compared with that in hPSC-derived hepatoblast-like cells. Three genes were selected as predictors of low propensity for hepatic differentiation. Expression of these genes was investigated in 23 hPSC clones. Review of representative cells by induction of hepatic differentiation suggested that low prediction scores were linked with low hepatic differentiation. Thus, our model using gene expression ranking and bioinformatic analysis could reasonably predict poor differentiation propensity of hPSC lines.
源自人类多能干细胞（hPSC）的功能性肝细胞有潜力成为药物开发早期阶段预测药物性肝毒性的工具。然而，据报道hPSC系向特定谱系分化的倾向有所不同。预测hPSC向肝细胞分化的低倾向能力将有助于选择有用的hPSC克隆，并显著加速用于药物研究的hPSC衍生肝细胞的开发。在本研究中，我们比较了五个hPSC系中与肝分化相关基因的表达，其中包括已知可分化为肝细胞的人类胚胎干细胞系H9，以及一个据报道肝分化倾向较差的hPSC系。通过双向聚类分析得出区分未分化hPSC、hPSC衍生的肝母细胞样分化细胞和原代人肝细胞的基因。比较了未分化hPSC中基因的表达水平顺序与hPSC衍生的肝母细胞样细胞中的表达水平顺序。选择了三个基因作为肝分化低倾向的预测指标。在23个hPSC克隆中研究了这些基因的表达。通过诱导肝分化对代表性细胞进行评估表明，低预测分数与低肝分化相关。因此，我们使用基因表达排名和生物信息学分析的模型可以合理地预测hPSC系的低分化倾向。
