Transplantation of human adipose stem cell-derived hepatocyte-like cells with restricted localization to liver using acellular amniotic membrane

Abstract

IntroductionAdult stem cell-derived hepatocytes transplantation holds considerable promise for future clinical individualized therapy of liver failure or dysfunction. However, the low engraftment of the available hepatocytes in the liver disease microenvironment has been a major obstacle.MethodsAcellular human amniotic membrane was developed as a three-dimensional scaffold and combined with hepatocyte-like cells derived from human adipose stem cells to engineer a hepatic tissue graft that would allow hepatocyte engraftment in the liver effectively.ResultsThe hepatic tissue grafts maintained hepatocyte-specific gene expression and functionality in vitro. When transplanted into the surgical incision in livers for engraftment, the engineered hepatic grafts significantly decreased the degree of liver injury caused by a carbon tetrachloride treatment and generated cords that were similar to the ductal plates in the liver between the acellular human amniotic membrane and the liver of receipts at day 3 post-transplantation. The hepatic tissue grafts maintained the expression of human hepatocyte-specific markers albumin, hepatocyte nuclear factor 4α, and cytochrome P450 2B6 in the liver of receipts, and acquired human-specific drug metabolism ability at eight weeks post-transplantation.ConclusionsThe acellular human amniotic membrane has the ability to maintain the functional phenotype of the hepatocyte-like cells derived from human adipose stem cells. Functional acellular human amniotic membrane-hepatocytes grafts integrated with the liver decreases the acute liver injury of mice. These engineered tissue constructs may support stem cell-based individualized therapy for liver disease and for bioartificial liver establishment.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0208-9) contains supplementary material, which is available to authorized users.

Endothelial cell‐derived matrix promotes the metabolic functional maturation of hepatocyte via integrin‐Src signalling

“…SEM analysis was performed as previously described . The samples were examined using a Hitachi S‐4800 scanning electron microscope (Hitachi, Tokyo, Japan).…”

“…The severe shortage of liver donors for human primary adult hepatocyte has prompted in vitro efforts to produce the functional hepatocyte‐like cells (HLCs) from stem cells . Human adipose stem cells (hASCs) have the same genetic makeup as the donor patient, making them an ideal cell source for disease modelling, hepatotoxicity testing, and cell‐based therapies . As a result, HLCs have been differentiated from hASCs in vitro by toning soluble maintenance factors or microRNA to direct cell fate .…”

“…However, the ability of stem cell‐derived HLCs to replicate the function of endogenous cells is limited by a blunted phenotype with reduced metabolic enzyme activity . The key challenge is to recreate cell–their surrounding extracellular matrix (ECM) interactions, that is in vivo liver microenvironment, which plays a vital role during liver development, and hepatocyte functional maturity . The ECM proteins produced provide not only an adhesive substrate for integrins but also a platform for the transduction of intracellular signalling events that regulate a host of cell functions and that play an important role in maintaining the differentiated phenotype .…”

“…Recent studies have suggested that combinatorial biochemical ECM signalling and acellular tissue‐derived matrix are more effective than traditional collagen sandwich cultures to functional maturation of HLCs. Considering the complexity of the ECM in vivo environment, it is difficult to elucidate the comprehensive roles of the naive ECM in hepatocyte differentiation using a mixture of single ECM molecules.…”

Mesenchymal stem cells seeded on a human amniotic membrane improve liver regeneration and mouse survival after extended hepatectomy

“…A tissue‐engineered product that combines MSCs and acellular HAM has recently been characterized (Roux et al, ). Moreover, a 3D transplantable hepatic graft combining HAM and hepatocyte‐like cells has been used for the treatment of carbon tetrachloride‐induced acute liver injury in mice (Yuan et al, ).…”