In Vitro Hepatocyte Differentiation of Mesenchymal Stem Cells Derived from Human Bone Marrow

Abstract

Introduction : In addition to long-term self-renewal capacity, human mesenchymal stem cells(MSCs) possess versatile differentiation potential ranging from mesenchyme-related multipotency to neuroectodermal and endodermal competency. Of particular concern is hepatogenic potential that can be used for liver-directed stem cell therapy and transplantation. In this study examined whether human bone marrow-derived MSCs are able to differentiate into hepatocytes. Materials and methods : MSCs were cultured 2-step protocol with use of hepatocyte growth factor and oncostatin M. In the course of cell differentiation, cell morphology was observed by LM, EM and PLD activity and the expression of ALB, AFP, CK- 18, CK-19, CPS, GS and GAPDH of heaptocyte were confirmed by Western blot analysis and RT-PCR. Hepatocyte functional activity were confirmed by glycogen storage and uptake of low-density lipoprotein(LDL). Results : After 3 weeks of induction, cuboidal morphology, which is characteristic of hepatocytes, was observed, and also cell expressed marker protein specific of liver cell such as albumin, CK 18, and PEPCK. The presence of stored glycogen, as determined by PAS staining, was visualized at 4 weeks differentiation. After 6 weeks of differentiation, hepatocytes demonstrated the ability to uptake significant levels of LDL. Early phase of differentiation, we observed morphologic change and cell organelles including Golgi body, mitochondria and ER by EM. Also PLD activity in the hepatic differentiation cells increased twofold or more at the 30 min point. Conclusions : MSCs from human bone marrow could differentiate into hepatocyte or hepatocyte-like cells in the differentiation media including HGF, FGF, EGF and OSM. Based on these observation, we conclude that human MSCs retain hepatogenic potential suitable for cell therapy and transplantation against intractable liver diseases. 1