Background. Mesenchymal stem cells (MSCs), isolated from bone marrow, adipose tissue, and umbilical cord tissue, have been known to differentiate into hepatocyte-like cells. MSCs can also be easily obtained from umbilical cord lining membrane (CLMSCs). CLMSCs are more primitive MSCs than those isolated from other tissue sources. Objectives. The aim of this study was to investigate the in vitro differentiation of CLMSCs into hepatocyte lineage. Materials and Methods. In this study, CLMSCs were isolated through a tissue attachment method. Cells were characterized for expression of MSC-specific markers and differentiation potency. CLMSCs were induced to differentiate into hepatocytes by a simple two-step protocol. Differentiated cells were examined for the expression of hepatocyte-specific markers and hepatocyte functions. Results. CLMSCs expressed MSC-specific markers and differentiated into adipocytes and osteoblasts. RT-PCR, real-time qRT-PCR, Western blot, and immunocytochemistry analyses demonstrated that differentiated CLMSCs, having hepatocyte-like morphology, expressed several liver-specific markers, such as ALB, AFP, CK18, and CK19, at both mRNA and protein levels following hepatocyte differentiation. Furthermore, periodic acid-Schiff staining and low-density lipoprotein (LDL) uptake assay showed that differentiated cells could store glycogen and uptake LDL. Conclusion. This study demonstrated that CLMSCs can differentiate into functional hepatocyte-like cells. CLMSCs can serve as a favorable cell source for tissue engineering in the treatment of liver disease. 1. Introduction Mesenchymal stem cells (MSCs) are referred to as highly proliferating and adherent fibroblastic cells featuring a unique expression profile of cell surface molecules. Like any other stem cells, they own capacity of self-renewal and multilineage differentiation [1]. MSCs can be obtained from various tissue sources, such as bone marrow (BM), adipose tissue (AD), and umbilical cord (UC) [2, 3]. BM is the main source for MSC isolation and BM-derived MSC (BMMSCs) have been extensively studied. However, the disadvantages of autologous BMMSCs, such as a highly invasive procedure for the donors and a significant decrease in both quantity and differentiation potential of cells with age, have restricted their use in tissue engineering and organogenesis [4]. Hence, it has accelerated the search for alternative sources of stem cells. In recent years, many research groups have focused on various types of umbilical cord-derived MSCs (UCMSCs), including UC matrix stem cells, UC
References
[1]
G. Chamberlain, J. Fox, B. Ashton, and J. Middleton, “Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing,” Stem Cells, vol. 25, no. 11, pp. 2739–2749, 2007.
[2]
W. Wagner, F. Wein, A. Seckinger et al., “Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood,” Experimental Hematology, vol. 33, no. 11, pp. 1402–1416, 2005.
[3]
D. Baksh, R. Yao, and R. S. Tuan, “Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow,” Stem Cells, vol. 25, no. 6, pp. 1384–1392, 2007.
[4]
K. Stenderup, J. Justesen, C. Clausen, and M. Kassem, “Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells,” Bone, vol. 33, no. 6, pp. 919–926, 2003.
[5]
J. Dalous, J. Larghero, and O. Baud, “Transplantation of umbilical cord-derived mesenchymal stem cells as a novel strategy to protect the central nervous system: technical aspects, preclinical studies, and clinical perspectives,” Pediatric Research, vol. 7, no. 4, pp. 482–490, 2012.
[6]
R. Friedman, M. Betancur, L. Boissel, H. Tuncer, C. Cetrulo, and H. Klingemann, “Umbilical cord mesenchymal stem cells: adjuvants for human cell transplantation,” Biology of Blood and Marrow Transplantation, vol. 13, no. 12, pp. 1477–1486, 2007.
[7]
M. Secco, E. Zucconi, N. M. Vieira et al., “Multipotent stem cells from umbilical cord: cord is richer than blood!,” Stem Cells, vol. 26, no. 1, pp. 146–150, 2008.
[8]
M. J. Seo, S. Y. Suh, Y. C. Bae, and J. S. Jung, “Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo,” Biochemical and Biophysical Research Communications, vol. 328, no. 1, pp. 258–264, 2005.
[9]
I. J. Fox and J. R. Chowdhury, “Hepatocyte transplantation,” American Journal of Transplantation, vol. 4, supplement 6, pp. 7–13, 2004.
[10]
S. C. Strom, R. A. Fisher, M. T. Thompson et al., “Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure,” Transplantation, vol. 63, no. 4, pp. 559–569, 1997.
[11]
P. A. Lysy, D. Campard, F. Smets, M. Najimi, and E. M. Sokal, “Stem cells for liver tissue repair: current knowledge and perspectives,” World Journal of Gastroenterology, vol. 14, no. 6, pp. 864–875, 2008.
[12]
S. H. Hong, E. J. Gang, J. A. Jeong et al., “In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells,” Biochemical and Biophysical Research Communications, vol. 330, no. 4, pp. 1153–1161, 2005.
[13]
R. Taléns-Visconti, A. Bonora, R. Jover et al., “Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells,” World Journal of Gastroenterology, vol. 12, no. 36, pp. 5834–5845, 2006.
[14]
H.-H. Yoon, B.-Y. Jung, Y.-K. Seo, K.-Y. Song, and J.-K. Park, “In vitro hepatic differentiation of umbilical cord-derived mesenchymal stem cell,” Process Biochemistry, vol. 45, no. 12, pp. 1857–1864, 2010.
[15]
S. Snykers, T. Vanhaecke, P. Papeleu et al., “Sequential exposure to cytokines reflecting embryogenesis: the key for in vitro differentiation of adult bone marrow stem cells into functional hepatocyte-like cells,” Toxicological Sciences, vol. 94, no. 2, pp. 330–341, 2006.
[16]
H. Kanazawa, Y. Fujimoto, T. Teratani et al., “Bone marrow-derived mesenchymal stem cells ameliorate hepatic ischemia reperfusion injury in a rat model,” PLoS ONE, vol. 6, no. 4, Article ID e19195, 2011.
[17]
M. Mouiseddine, S. Fran？ois, M. Souidi, and A. Chapel, “Intravenous human mesenchymal stem cells transplantation in NOD/SCID mice preserve liver integrity of irradiation damage,” Methods in Molecular Biology, vol. 826, pp. 179–188, 2012.
[18]
K. Kita, G. G. Gauglitz, T. T. Phan, D. N. Herndon, and M. G. Jeschke, “Isolation and characterization of mesenchymal stem cells from the sub-amniotic human umbilical cord lining membrane,” Stem Cells and Development, vol. 19, no. 4, pp. 491–501, 2010.
[19]
K. J. Livak and T. D. Schmittgen, “Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method,” Methods, vol. 25, no. 4, pp. 402–408, 2001.
[20]
M. Dominici, K. le Blanc, I. Mueller et al., “Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement,” Cytotherapy, vol. 8, no. 4, pp. 315–317, 2006.
[21]
C. Schmidt, F. Bladt, S. Goedecke et al., “Scatter factor/hepatocyte growth factor is essential for liver development,” Nature, vol. 373, no. 6516, pp. 699–702, 1995.
[22]
J. Jung, M. Zheng, M. Goldfarb, and K. S. Zaret, “Initiation of mammalian liver development from endoderm by fibroblast growth factors,” Science, vol. 284, no. 5422, pp. 1998–2003, 1999.
[23]
Y. Sakai, J. Jiang, N. Kojima, T. Kinoshita, and A. Miyajima, “Enhanced in vitro maturation of fetal mouse liver cells with oncostatin M, nicotinamide, and dimethyl sulfoxide,” Cell Transplantation, vol. 11, no. 5, pp. 435–441, 2002.
[24]
G. K. Michalopoulos, W. C. Bowen, K. Mule, and J. Luo, “HGF-, EGF-, and dexamethasone-induced gene expression patterns during formation of tissue in hepatic organoid cultures,” Gene Expression, vol. 11, no. 2, pp. 55–75, 2003.
[25]
E. Schmelzer, E. Wauthier, and L. M. Reid, “The phenotypes of pluripotent human hepatic progenitors,” Stem Cells, vol. 24, no. 8, pp. 1852–1858, 2006.
