Laminin matrix promotes hepatogenic terminal differentiation of human bone marrow mesenchymal stem cells


1 Animal and Marine Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

2 Department of Clinical Biochemistry , Faculty of Medical Sciences ,Tarbiat Modares University , Tehran, Iran

3 Biomaterials and Tissue Engineering Department, Stem Cell Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran


Objective(s):The application of stem cells holds great promises in cell transplants. Considering the lack of optimal in vitro model for hepatogenic differentiation, this study was designed to examine the effects of laminin matrix on the improvement of in vitro differentiation of human bone marrow mesenchymal stem cells (hBM-MSC) into the more functional hepatocyte-like cells.
Materials and Methods:Characterization of the hBM-MSCs was performed by immunophenotyping and their differentiation into the mesenchymal-derived lineage. Then, cells were seeded on the laminin-coated or tissue culture polystyrene (TCPS). The differentiation was carried out during two steps. Afterward, the expression of hepatocyte markers such as AFP, ALB, CK-18, and CK-19 as well as the expression of C-MET, the secretion of urea, and the activity of CYP3A4 enzyme were determined. Moreover, the cytoplasmic glycogen storage was examined by periodic acid–Schiff (PAS) staining.
Results:The results demonstrated that the culture of hBM-MSC on laminin considerably improved hepatogenic differentiation compared to TCP group. A significant elevated level of urea biosynthesis and CYP3A4 enzyme activity was observed in the media of the laminin-coated differentiated cells (P<0.05). Furthermore higher expressions of both AFP and ALB were determined in cells differentiated on laminin matrix. Glycogen accumulation was not detected in the undifferentiated hBM-MSCs, however, both differentiated cells in laminin and TCPS groups demonstrated the intracellular glycogen accumulation on day 21 of hepatogenic differentiation.
Conclusion:Taken together, these findings may indicate that laminin matrix can improve terminal differentiation of hepatocyte-like cells from hBM-MSCs. Thus, laminin might be considered as a suitable coating in hepatic tissue engineering designs.


1. Li J, Li M, Niu B, Gong J. Therapeutic potential of stem cell in liver regeneration. Front Med 2011; 5:26-32.

2.O'Leary JG, Lepe R, Davis GL. Indications for liver transplantation. Gastroenterology 2008; 134:1764-1776.

3.Kosmacheva S, Seviaryn I, Goncharova N, Petyovka N, Potapnev M. Hepatogenic potential of human bone marrow and umbilical cord blood mesenchymal stem cells. Bull Exp Biol Med 2011; 151:142-149.

4.Gómez‐Lechón MJ, Jover R, Donato T, Ponsoda X, Rodriguez C, Stenzel KG, et al. Long‐term expression of differentiated functions in hepatocytes cultured in three‐dimensional collagen matrix. J Cell Physiol 1998; 177:553-562.

5.Vestentoft PS. Development and molecular composition of the hepatic progenitor cell niche. Dan Med J 2013; 60:B4640.

6.Spradling A, Drummond-Barbosa D, Kai T. Stem cells find their niche. Nature 2001; 414:98-104.

7.Fuchs E, Tumbar T, Guasch G. Socializing with the neighbors: stem cells and their niche. Cell 2004; 116:769-778.

8.Scadden DT. The stem-cell niche as an entity of action. Nature 2006; 441:1075-1079.

9.Schaffner F, Poper H. Capillarization of hepatic sinusoids in man. Gastroenterology 1963; 44:239-242.

10. Hahn E, Wick G, Pencev D, Timpl R. Distribution of basement membrane proteins in normal and fibrotic human liver: collagen type IV, laminin, and fibronectin. Gut 1980; 21:63-71.

11. He H, Liu X, Peng L, Gao Z, Ye Y, Su Y, et al. Promotion of hepatic differentiation of bone marrow mesenchymal stem cells on decellularized cell-deposited extracellular matrix. Biomed Res Int 2013; 2013.

12. Terada T, Nakanuma Y. Expression of tenascin, type IV collagen and laminin during human intrahepatic bile duct development and in intrahepatic cholangiocarcinoma. Histopathology 1994; 25:143-150.

13. Martinez-Hernandez A, Delgado FM, Amenta P. The extracellular matrix in hepatic regeneration. Localization of collagen types I, III, IV, laminin, and fibronectin. Lab Invest 1991; 64:157-166.

14. Wewer UM, Albrechtsen R. Carcinoma-associated perisinusoidal laminin may signal tumour cell metastasis to the liver. Virchows Arch A Pathol Anat Histopathol 1992; 421:87-93.

15. Quondamatteo F, Scharif K, Herken R. Changes in laminin immunoreactivity as a marker for the state of liver preservation. Histochem J 1994; 26:827-832.

16. Friedman SL, Roll FJ, Boyles J, Bissell DM. Hepatic lipocytes: the principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci USA 1985; 82:8681-8685.

17. Knittel T, Janneck T, Muller L, Fellmer P, Ramadori G. Transforming growth factor β1‐regulated gene expression of Ito cells. Hepatology 1996; 24:352-360.

18. Nikolova G, Strilic B, Lammert E. The vascular niche and its basement membrane. Trends Cell Biol 2007; 17:19-25.

19. Tanentzapf G, Devenport D, Godt D, Brown NH. Integrin-dependent anchoring of a stem-cell niche. Nat Cell Biol 2007; 9:1413-1418.

20. Tate MC, Garcı́a AJ, Keselowsky BG, Schumm MA, Archer DR, LaPlaca MC. Specific beta1 integrins mediate adhesion, migration, and differentiation of neural progenitors derived from the embryonic striatum. Mol Cell Neurosci 2004; 27:22-31.

21. Hayashi Y, Furue MK, Okamoto T, Ohnuma K, Myoishi Y, Fukuhara Y, et al. Integrins regulate mouse embryonic stem cell self‐renewal. Stem Cells 2007; 25:3005-3015.

22. Kazemnejad S, Allameh A, Soleimani M, Gharehbaghian A, Mohammadi Y, Amirizadeh N, et al. Biochemical and molecular characterization of hepatocyte-like cells derived from human bone marrow mesenchymal stem cells on a novel three-dimensional biocompatible nanofibrous scaffold. J Gastroenterol Hepatol 2009; 24:278-287.

23. Ayatollahi M, Soleimani M, Tabei SZ, Salmani MK. Hepatogenic differentiation of mesenchymal stem cells induced by insulin like growth factor-I. World J Stem Cells 2011; 3:113-121.

24. Kallis YN, Robson AJ, Fallowfield JA, Thomas HC, Alison MR, Wright NA, et al. Remodelling of extracellular matrix is a requirement for the hepatic progenitor cell response. Gut 2011; 60:525-533.

25. Takayama K, Nagamoto Y,  Mimura N, Tashiro K, Sakurai F, Tachibana M, et al. Long-Term Self-Renewal of Human ES/iPS-Derived Hepatoblast-like Cells on Human Laminin 111-Coated Dishes. Stem Cell Reports 2013; 1:322-335.