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.

Mesenchymal stem cells accelerate liver regeneration in acute liver failure animal model

“…Specifically, the regeneration process is induced by the activation of liver progenitor cells (LPCs) derived from endogenous progenitors (local cells) and or bone marrowderived circulating cells, particularly mesenchymal stem cells (MSCs) 3 . Studies have demonstrated that MSCs have high differentiation potential to specific cells, including primary hepatocytes 4 . These facts suggest that MSCs could accelerate the regeneration process of ALF.…”

Different approaches for transformation of mesenchymal stem cells into hepatocyte-like cells

“…Finally, they showed that the differentiated cells can improve liver injury in an animal model. In their studies, hMSCs were directly converted into hepatocytes using a seven-miRNA combination and they claimed that in comparison with conventional methods, the seven-miRNA combination promoted hepatic differentiation of hMSCs more quickly and efficiently [79,80]. In another study, Khosravi et al used miR-106a, miR-574-3p, and miR-451 for hepatic differentiation for in vitro UC-MSCs and confirmed the production of ALB [81].…”

Alternative Cell Sources for Liver Parenchyma Repopulation: Where Do We Stand?

“…Since then, a large number of studies tested the possibility of repopulating the liver with in vitro hepatic differentiated MSCs. To generate hepatocytes from MSC, a multitude of approaches has been used including the addition of cytokine cocktails [65,[86][87][88][89][90][91][92][93][94][95], small molecules [96], and/or microRNAs (miRNAs) [97][98][99] to the cell culture medium and recreation of the physicochemical characteristics of a hepatocyte niche micro-environment [100,101]. Generally, HLCs derived from MSCs possess some mature and fetal hepatocyte features as has been shown by transcriptome and functional studies [102,103].…”

“…In general, wild-type or immunodeficient mice without liver damage inducing transgene were used, and liver injury was caused by treatment with CCl 4 . A number of reports demonstrated presence of single cells or small groups of hALB positive cells throughout the liver (RI between 1 to 5%) 1 to 14 days after transplantation [86,89,97,98,100,105]. Most studies did not perform long-term follow-up analysis.…”