RNA-seq and scRNA-seq analysis of the efficient generation of functional human hepatocytes from chemically induced pluripotent stem cells

Here, we developed a new differentiation protocol by mimicking the two-stage development of hepatoblasts to potentiate their transition to hepatic progenitor cells (hiHPCs), which permits the high-efficiency generation of hiHPCs from chemically induced pluripotent stem cells (hCiPSCs). Moreover, hCiPSC-derived hiHPCs can further differentiate into mature hepatocytes (hiHEPs). We use single-cell RNA sequencing (scRNA-seq) to analyze the two-stage of hepatoblasts indicating the similarity between hepatoblasts differentiated in vitro and in vivo. In addition, we show that the global gene expression profile of hiHPCs was indistinguishable from that of human fetal liver cells (hFLCs) and hCiPSC-derived hiHEPs resembled F-PHHs by using RNA sequencing (RNA-seq). To dissect the two-stage transition of hepatoblasts, the process was analyzed by scRNA-seq. Moreover, hCiPSC-derived hiHPCs and hiHEPs are similar to hFLCs and F-PHHs respectively by RNA-seq.