title: Induction of hepatocyte-like cells from human umbilical cord-derived mesenchymal stem cells by defined microRNAs.
abstract: Generating functional hepatocyte-like cells (HLCs) from mesenchymal stem cells (MSCs) is of great urgency for bio-artificial liver support system (BALSS). Previously, we obtained HLCs from human umbilical cord-derived MSCs by overexpressing seven microRNAs (HLC-7) and characterized their liver functions in vitro and in vivo. Here, we aimed to screen out the optimal miRNA candidates for hepatic differentiation. We sequentially removed individual miRNAs from the pool and examined the effect of transfection with remainder using RT-PCR, periodic acid-Schiff (PAS) staining and low-density lipoprotein (LDL) uptake assays and by assessing their function in liver injury models. Surprisingly, miR-30a and miR-1290 were dispensable for hepatic differentiation. The remaining five miRNAs (miR-122, miR-148a, miR-424, miR-542-5p and miR-1246) are essential for this process, because omitting any one from the five-miRNA combination prevented hepatic trans-differentiation. We found that HLCs trans-differentiated from five microRNAs (HLC-5) expressed high level of hepatic markers and functioned similar to hepatocytes. Intravenous transplantation of HLC-5 into nude mice with CCl4-induced fulminant liver failure and acute liver injury not only improved serum parameters and their liver histology, but also improved survival rate of mice in severe hepatic failure. These data indicated that HLC-5 functioned similar to HLC-7 in vitro and in vivo, which have been shown to resemble hepatocytes. Instead of using seven-miRNA combination, a simplified five-miRNA combination can be used to obtain functional HLCs in only 7 days. Our study demonstrated an optimized and efficient method for generating functional MSC-derived HLCs that may serve as an attractive cell alternative for BALSS.
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Cui
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Xinmin
authors.initials: X
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Guanya
authors.initials: G
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Hou
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Weile
authors.initials: W
authors.lastname: Cai
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Zheyi
authors.initials: Z
authors.lastname: Han
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Han
mesh_terms: D000818:Animals; D002251:Carbon Tetrachloride; D054338:Cell Transdifferentiation / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D006801:Humans; D017093:Liver Failure / Q000235:genetics / Q000473:pathology / Q000628:therapy; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D016019:Survival Analysis; D014162:Transfection; D014470:Umbilical Cord / Q000166:cytology*
chemical_list: D035683:MicroRNAs; D002251:Carbon Tetrachloride
keywords: BALSS; CCl4; hepatic differentiation; mesenchymal stem cells; miRNA