Z Gastroenterol 2014; 52 - P_4_22
DOI: 10.1055/s-0033-1360975

Hepatogenic differentiated murine mesenchymal stem cells display functional qualities of perivenous and/or periportal hepatocytes

S Winkler 1, S Brückner 1, P Stock 1, M Hempel 1, B Christ 1
  • 1University of Leipzig, Department of Visceral, Transplantation, Thoracic and Vascular Surgery, Leipzig, Germany

Background: Mesenchymal stem cells possess the ability to differentiate into hepatocyte-like cells in vitro. Thus, they open the possibility for use as therapeutic alternative instead of hepatocyte or even whole organ transplantation in different chronic (metabolic) liver diseases such as e.g. alpha-1-antitrypsin deficiency. This study aimed to examine functional parameters of murine mesenchymal stem cells after hepatogenic differentiation in order to develop a pre-clinical assessment of their potential for clinical application in chronic liver disease models.

Methods: Mesenchymal stem cells were isolated from murine adipose tissue of immunodeficient Pfp/Rag2-/- mice and were cultured in differentiation media after reaching 90% confluency. After 0, 7, 14 and 21 days, hepatogenic features like glycogen storage (PAS-staining), urea synthesis and the enzyme activity of ethoxyresorufin-O-deethylase (EROD) were measured. Periportal markers like phosphoenolpyruvatcarboxykinase (PCK1) and perivenous markers like glutamine synthase were stained immunocytochemically.

Results: The procedure of differentiation induced morphological changes from a fibroblastoid structure of undifferentiated cells into a polygonal shape of differentiated cells. Glycogen deposits were found from day 7 of differentiation further increasing until day 21. After 14 days of differentiation, functional markers like urea synthesis and EROD enzyme activity were significantly increased. Sub-populations of cells stained either positive for PCK1 or GS or both as identified by immunocytochemistry.

Conclusion: Hepatogenic differentiated murine stem cells feature specific perivenous and periportal hepatocyte functions already after 14 and 21 days of differentiation. Transplantation experiments using subpopulations of differentiated cells in a mouse model of chronic alpha-1-antitrypsin deficiency will provide more insight into their therapeutic potential.