Abstract The differentiation of human induced pluripotent stem cells (hiPSCs) into functional hepatocytes has the potential to solve the shortage of human primary liver cells and would be of use in drug screening. In this frame, we developed a hiPSCs maturation strategy in microfluidic biochips, using a liver-on-chip approach, a promising technology mimicking in vivo physiology. Hepato-like tissues differentiated in biochips presented advanced liver features, including ALB and CYP3A4 expressing cells. The metabolomics and transcriptomics profiles of hepato-likes cells differentiated either in biochips or Petri dishes were integrated to compare their functionalities. The multi-omics analysis revealed 41 metabolites and 302 genes differentially expressed. Overall, biochip environment lead to higher degree of hepatic differentiation demonstrated by an increase in the metabolic production of lipids, fatty acids and biliary acids, which was confirmed at the transcriptome level by the modulation of expression for genes involved in related signaling pathways. This observation was correlated with higher production of fructose in biochips, together with down-regulation of genes engaged in glycolysis. In parallel, increased activity of the Krebs cycle, pentose phosphate shuttle, and fatty acid beta oxidation was observed in tissues cultured in Petri. Besides, the modulation of nitrogen metabolism was observed in transcriptomic data and confirmed by an intense production of glutamine, putrescine and creatinine and by the higher consumption of spermidine measured in Petri.

中文翻译：

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hiPSCs 衍生肝细胞在微流体环境中的代谢组学和转录组学特征的整合

摘要 将人类诱导多能干细胞（hiPSCs）分化为功能性肝细胞有可能解决人类原代肝细胞短缺的问题，并将用于药物筛选。在这个框架中，我们使用肝脏芯片方法开发了微流控生物芯片中的 hiPSCs 成熟策略，这是一种模拟体内生理学的有前途的技术。在生物芯片中分化的肝样组织呈现出先进的肝脏特征，包括表达 ALB 和 CYP3A4 的细胞。整合了在生物芯片或培养皿中分化的肝样细胞的代谢组学和转录组学特征，以比较它们的功能。多组学分析揭示了 41 个代谢物和 302 个差异表达的基因。全面的，生物芯片环境导致更高程度的肝脏分化，脂质、脂肪酸和胆汁酸的代谢产生增加，这在转录组水平通过相关信号通路相关基因表达的调节得到证实。这一观察结果与生物芯片中更高的果糖产量以及参与糖酵解的基因下调有关。同时，在 Petri 培养的组织中观察到三羧酸循环、磷酸戊糖穿梭和脂肪酸 β 氧化的活性增加。此外，在转录组数据中观察到氮代谢的调节，并通过谷氨酰胺、腐胺和肌酐的大量产生以及在 Petri 中测量到的亚精胺的更高消耗来证实。