Differentiation of Rat Bone Marrow Mesenchymal Stem Cell into Hepatocyte-Like Cells in Natural Scaffolds and Transplantation into Liver-damage Rat

Nasiri Z *, Soleimani M , Pedram M , Sasani F , Mosavi Z ,


Objective: Tissue engineering is a promising approach to developing hepatic tissue suitable for the functional replacement of a failing liver .previous study have been showed that Bone marrow mesenchymal stem cells (BMSc) have plasticity to differentiate into hepatocyte-like in monolayer . The aim of the present study was to investigate in vitro study was to evaluate the morphology and hepatocyte differentiation of bone marrow derived mesenchymal stem cells with in natural scaffold that could influence the proliferation rate and survival of rat hepatocytes both during long term culture and after in vivo transplantation. Materials and Methods: In the present study, BMSCs was isolated from rat male SD (3-4 weeks) Marrow-derived hepatocyte were seeded into porous natural coral scaffolds in a density of 1 × 106/mL in 300 µL cell suspension, cell cultured were treated with HGF, EGF, Dexamethasone and OSM in two step protocol and an animal model of engraftment in Fibrosis liver induced using carbon tetrachloride (CCl4) in rats .after 14 day's tissue-engineered hepatocyte valves were analyzed by RT-PCR and scanning electron microscopy Proliferation of the seeded cells on the scaffolds was detected using the MTT assay. After 8-10 wk of MSCs administration, blood samples were collected to measure the albumin, SGOT, SGPT concentration all rats were killed and fibrosis index were assessed by histopathology. Results: SEM micrographs of cells showed that cells adhered and proliferated well on the outer and inner surfaces of these natural scaffolds the result of immunofluorescent analysis revealed the expression of albumin (ALB) and alpha feto protein (AFP) and anti –hepatocyte from day 7 to day 28 similar the finding of expression of Alb ,CK19 AFP,…mRNA by RT-PCR. In vivo results demonstrated the biocompatibility of natural scaffold implanted in rat fibrosis and significantly increased the serum albumin concentration and vascularization of porous scaffolds implanted on liver lobes and improved hepatocyte engraftment. Conclusion: All these results indicate that rMSCs differentiated into hepatocyte in natural scaffold, Implantable engineered liver tissue using natural coral as scaffold for growth and differentiation of hepatocytes has been developed. This technique is an attractive tool for the development of liver tissue engineering and may provide a source of differentiated cells for treatment of liver diseases.