Four Step Differentiation of Umbilical Cord Derived Mesenchymal Stem Cells Within Hydrogel Scaffold Into Hepatocytes

Due to increasing demand for liver tissue engineering, three-dimensional (3D) liver cells culture techniques have been proposed. Therefore, the aim of the present study was to examine the cells isolation and expansion of umbilical cord derived mesenchymal stem cells and in vitro 2D and 3D hepatocyte differentiation. Also functional characteristics of hepatocytes were analyzed. The study performed in several phases. In the first umbilical cord derived mesenchymal stem cells obtained and isolated, thereafter cellss expanded. Determination of Immunophenotype using Flowcytometry performed by DAKO – Galaxy Hepatic differentiation UC-MSCs was performed by four step sequential method using FGF-4, ITS, HGF, dexamethasone, glucagon, OSM and TSA. Urea production was quantified by ELISA. Section of tissue constructs stained with hematoxyllin and eosin for histological examination. MSCs isolated from umbilical cord expressed mesenchymal surface antigen such as CD73, but were negative against CD31. Several cell clusters mainly between the round cells were observed in alginate scaffold after 3d differentiation. Urea production was increased time- dependable and was significantly higher in the experimental group of 3D culture (P=0.001). Tissue construct of 3D culture revealed multicellular tissue with several euchromatin cell plates. The finding of the present study indicated that four step differentiation of umbilical cord derived mesenchymal stem cells within hydrogel scaffold induced functionally and morphologically characteristics of hepatocytes such as urea production and cell plates.