Probes were ranked by the weighted average difference method?(Kadota et al., 2008), and the top 5% (2445 probes) were defined as differentially expressed genes. Supplementary file 4: Significantly enriched gene sets (Nom p 0.05) in Hep-i(+) cells compared with Hep-i(-) cells (assessed by GSEA). elife-47313-supp4.xlsx (13K) DOI:?10.7554/eLife.47313.030 Supplementary file 5: Significantly enriched gene sets (Nom p 0.05) in Hep-i(-) cells compared with Hep-i(+) cells (assessed by GSEA). elife-47313-supp5.xlsx (16K) DOI:?10.7554/eLife.47313.031 Supplementary file 6: Significantly enriched (NOM p 0.05) gene sets in hCLiP-chimera-derived hepatocytes in comparison with PHHs. elife-47313-supp6.xlsx (18K) DOI:?10.7554/eLife.47313.032 Transparent reporting form. elife-47313-transrepform.docx (245K) DOI:?10.7554/eLife.47313.033 Data Availability StatementMicroarray transcriptome data are available with accession numbers “type”:”entrez-geo”,”attrs”:”text”:”GSE133776″,”term_id”:”133776″GSE133776 (Reprogramming of primary human hepatocytes (PHHs) into hCLiPs); “type”:”entrez-geo”,”attrs”:”text”:”GSE133777″,”term_id”:”133777″GSE133777 (Hepatic induction of hCLiPs); “type”:”entrez-geo”,”attrs”:”text”:”GSE133778″,”term_id”:”133778″GSE133778 (Characterization of long term-cultured of hCLiPs); “type”:”entrez-geo”,”attrs”:”text”:”GSE133779″,”term_id”:”133779″GSE133779 (Transcriptomic analysis of PHHs isolated from hCLiP-transplanted mouse chimeric liver). “type”:”entrez-geo”,”attrs”:”text”:”GSE133776″,”term_id”:”133776″GSE133776-“type”:”entrez-geo”,”attrs”:”text”:”GSE133779″,”term_id”:”133779″GSE133779 are included in Superseries “type”:”entrez-geo”,”attrs”:”text”:”GSE133797″,”term_id”:”133797″GSE133797. Comparative analysis of IPHH and APHH transcriptome is available with an accession number “type”:”entrez-geo”,”attrs”:”text”:”GSE134672″,”term_id”:”134672″GSE134672. The following datasets were generated: Takeshi Katsuda, Takahiro Ochiya. 2019. Reprogramming of primary human hepatocytes (PHHs) into hCLiPs. NCBI Gene Expression Omnibus. GSE133776 Takeshi Katsuda, Takahiro Ochiya. 2019. Hepatic induction of hCLiPs. NCBI Gene Expression Omnibus. GSE133777 Takeshi Katsuda, Takahiro Ochiya. 2019. Characterization of long term-cultured of hCLiPs. NCBI Gene Expression Omnibus. GSE133778 Takeshi Katsuda, Takahiro Ochiya. 2019. Transcriptomic analysis of PHHs Stattic isolated from hCLiP-transplanted mouse chimeric liver. NCBI Gene Expression Omnibus. GSE133779 Takeshi Katsuda, Takahiro Ochiya. 2019. Comparison between infant and adult primary human hepatocytes (PHHs) in terms of their responsiveness to FAC (FBS + A83-01 + CHIR99021) NCBI Gene Expression Omnibus. GSE134672 Abstract Hepatocytes are regarded as the only effective cell source for cell transplantation to treat liver diseases; however, their availability is limited due to a donor shortage. Thus, a novel cell source must be developed. We recently reported that mature rodent hepatocytes can be reprogrammed into progenitor-like cells with a repopulative capacity using small molecule inhibitors. Here, we demonstrate that hepatic progenitor cells can be obtained from human infant hepatocytes using the same strategy. These cells, named human chemically induced liver progenitors (hCLiPs), had a significant repopulative capacity in injured mouse livers following transplantation. hCLiPs redifferentiated into mature hepatocytes in vitro upon treatment with hepatic maturation-inducing factors. These redifferentiated cells exhibited cytochrome P450 (CYP) enzymatic activities in response to CYP-inducing molecules and these activities were Stattic comparable with those in primary human hepatocytes. These findings will facilitate liver cell transplantation therapy and drug discovery studies. and and was affected not only by the presence of AC but also by the culture duration, suggesting that AC-induced expression of these genes during in vitro culture. By contrast, expression of and was maintained, but not increased, upon culture in the presence of AC. Gene signature enrichment analysis (GSEA) comparing cells cultured in the presence of FBS and those cultured in FAC demonstrated that the majority of gene sets enriched in the latter cells were related Mouse monoclonal to COX4I1 to hepatic function (Figure 2G, Supplementary file 1), suggesting that AC also helped to maintain the hepatocytic characteristics of cultured hepatocytes. Although cell-cycle-related gene sets were also identified by GSEA, their enrichment scores were relatively low (Figure 2figure supplement Stattic 3A, Supplementary file 1). This Stattic is likely because cell proliferation was also increased in part by culture in FBS alone. Indeed, proliferation-related gene sets were enriched both in cells cultured in FBS only and in FAC compared with D1 hepatocytes (Figure 2figure supplement 3B and C, Supplementary file 2, 3). In summary, two small molecules, AC, together with FBS, support the proliferation of hepatic epithelial cells with characteristics of both hepatocytes and LPCs/BECs. Comparison of IPHHs and APHHs in terms of their responsiveness to FAC To investigate the difference regarding the responsiveness to FAC of IPHHs and APHHs, we compared their transpcriptome by microarray analysis. Hierarchical clustering of the whole transcriptome demonstrated that IPHHs cultured in FAC for 7 or 14 days formed a cluster distinct from those cultured in FBS (Figure 3A). In contrast, APHHs cultured in FAC for 7 or 14 days were not clearly separated from those cultured in FBS. These results suggest that APHHs are less sensitive to AC than IPHHs. GSEA indicated that many of the signaling pathways enriched for IPHHs cultured Stattic in FAC for 7 days compared with APHHs were cell-cycle-related pathways (Figure 3figure supplements 1 and ?and2)2) (we avoided comparing cells at D14, because lot 187271 APHHs were severely contaminated with NPCs at D14, as shown.