Transplantation Of Hepatocyte-like Cells Research Articles

Generation and transplantation of hepatocytes-like cells using human origin hepatogenic serum for acute liver injury treatment.

Liver failure is a critical disease for which regenerative therapies are still being explored. The major limitation in the use of a clinical grade, viable cell-based therapy approach is the scarce availability of sufficient number of in-vitro differentiated hepatocyte-like cells (HLC) that can induce regeneration and ameliorate liver injury. Here, we report for the first time an approach to engineer HLCs using sera of hyperbilirubin patients that act as a reservoir of differentiation factor. Utilizing our humanized approach, mesenchymal stem cells (hMSC) derived from umbilical cord tissue were transdifferentiated into HLC using patient-derived serum along with dimethyl sulfoxide (DMSO). We studied the effects of serum on the proliferation, cell cycle analysis, and apoptosis of hMSC by various differentiation combinations. We optimized the hepatic transdifferentiation ability of hMSC with hyperbilirubin serum treatment for a period of 7 days. Assessment of HLC functionalities was shown by quantifying the HLC spent medium for albumin and urea secretions. Transplantation of HLC in an acute liver injury (ALI) rat model showed an effective improvement in the liver function and histological changes in the liver. The results of this study suggest that hMSC-derived HLC using humanized hepatogenic serum holds a promising potential for cell transplantation, as an efficient therapy modality for liver failure in humans.

Amelioration of Hyperbilirubinemia in Gunn Rats after Transplantation of Human Induced Pluripotent Stem Cell-Derived Hepatocytes

SummaryHepatocyte transplantation has the potential to cure inherited liver diseases, but its application is impeded by a scarcity of donor livers. Therefore, we explored whether transplantation of hepatocyte-like cells (iHeps) differentiated from human induced pluripotent stem cells (iPSCs) could ameliorate inherited liver diseases. iPSCs reprogrammed from human skin fibroblasts were differentiated to iHeps, which were transplanted into livers of uridinediphosphoglucuronate glucuronosyltransferase-1 (UGT1A1)-deficient Gunn rats, a model of Crigler-Najjar syndrome 1 (CN1), where elevated unconjugated bilirubin causes brain injury and death. To promote iHep proliferation, 30% of the recipient liver was X-irradiated before transplantation, and hepatocyte growth factor was expressed. After transplantation, UGT1A1+ iHep clusters constituted 2.5%–7.5% of the preconditioned liver lobe. A decline of serum bilirubin by 30%–60% and biliary excretion of bilirubin glucuronides indicated that transplanted iHeps expressed UGT1A1 activity, a postnatal function of hepatocytes. Therefore, iHeps warrant further exploration as a renewable source of hepatocytes for treating inherited liver diseases.

Transplantation of Hepatocytelike Cells Derived from Umbilical Cord Stromal Mesenchymal Stem Cells to Treat Acute Liver Failure Rat

Background: Liver transplantation is the ultimate treatment for liver failure. However, its use is complicated by shortage of donor organs and numerous other problems. Thus, other methods for the treatment of liver failure, including cell therapy, are being considered. We have evaluated the capacity of hepatocyte-like cells differentiated from Wharton’s jelly mesenchymal stem cells (WJ-MSCs) to prevent or ameliorate acute liver failure in a rat model. Methods: WJ-MSCs were isolated from human umbilical cords and induced to differentiate into hepatocyte-like cells by incubation with a combination of liver-tissue culture medium and growth factors. The differentiated cells were evaluated with immunocytochemistry, RT-PCR, ammonia metabolism test, and glycogen storage for characteristics of functioning hepatocytes. The hepatocyte-like cells were transplanted into the portal vein of rats with acute liver failure induced by carbontetrachloride injection. Survival, serum liver function tests, and body weight of the animals were recorded. Livers of the rats 30 days after transplantation were harvested. Results: The differentiated hepatocyte-like cells had morphologic features and functional characteristics of mature hepatocytes: hepatocyte-related genes, conversion of ammonia to urea, albumin synthesis, glycogen storage, and declining values of alpha-fetoprotein. Transplantation of the hepatocyte-like cells into the rats with carbon tetrachloride-induced liver failure prevented death of the animals and prevented rising values of serum alanine transaminase (ALT), aspartate transaminase (AST), and bilirubin. Immunohistochemical study of rat liver tissues 30 days after transplantation revealed that the WJ-MSCs had survived and were producing albumin. Conclusions: WJ-MSCs of the human umbilical cord were could differentiated into hepatocyte-like cells in vitro. Transplantation of hepatocyte-like cells into the portal vein of rats with carbontetrachloride acute liver injury prevented death of the animals and irreversible liver injury. These findings may serve as a reference for future research in cell therapy of liver failure.