Hepatocyte Differentiation of Mesenchymal Stem Cells from Adipose Tissue of Immnunodeficient Mice

Abstract

Mesenchymal stem cells (MSCs) feature multipotent differentiation capacity including differentiation into hepatocyte-like cells in vitro and in vivo. Therefore, MSC represent a novel cell resource for the treatment of liver diseases or to overcome the shortage of organs for liver transplantation. The aim of this work was to isolate MSC from adipose tissue of immunodeficient mice as well as to verify their differentiation potential. Methods: MSCs were isolated from immunodeficient Pfp/Rag2−/− mice. They were cultured until they reached 90% confluency. After a demethylation step specialized growth- and differentiation conditions were chosen. After 0, 7, 14, and 21 days the morphology of the cells was documented microscopically. Mesenchymal (CD44, CD90, CD105) and haematopoetic (CD45, CD34) surface markers were determined by flow-cytometry. The gene expression of AFP as well as of hepatocyte markers like GS, HNF-4, Cyp3A11, PCK1 and albumin were measured by sq-RT-PCR. Results: The morphology of adult MSCs changed with progressing differentiation time from a spindle-shaped into a polygonal morphology. Mesenchymal surface markers were expressed at each time point and haematopoetic markers were hardly detectable after 21d of differentiation (1.6% of CD45 and 1.2% of CD34). The relative expression of all hepatic markers investigated increased continuously. APF was not expressed at any time point. Conclusion: Adult MSC from mouse adipose tissue may differentiate into hepatocyte-like cells displaying both morphological and functional hepatocyte features. The potential therapeutic benefit may now be analyzed in mouse models of acute and chronic liver diseases.