Ameliorating effect of encapsulated hepatocyte-like cells derived from umbilical cord in high mannuronic alginate scaffolds on acute liver failure in rats

Document Type : Original Article


1 Department of Anatomical Sciences, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

2 Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

3 Transplantation Ward, Ahvaz Golestan Hospital, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran


Objective(s): In this study, effects of encapsulated umbilical cord stem cells (UCSCs)-derived hepatocyte-like cells (HLCs) in high mannuronic alginate scaffolds was investigated on CCl4-induced acute liver failure (ALF) in rats.
Material and Methods: UCSCs were encapsulated in high mannuronic alginate scaffolds. Then the UCSCs differentiated into HLCs for treatment of CCl4-induced ALF in rats. Thirty rats randomly divided into 5 groups: Intoxicated group received only CCl4 to induce ALF. In other groups including cell-free, UCSCs and HLCs, alginate scaffolds were transplanted into the liver 4 days after CCl4 injection. Biochemical markers including albumin (ALB), blood urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated. Histological changes and gene expression of ALB, alpha-fetoprotein (AFP), and cytokeratin 18 (CK-18) were also assessed.
Results:  Expression of CK-18 significantly increased in HLCs compared to the UCSCs in vitro. This indicates that UCSCs can effectively differentiate into the HLCs. In CCl4-intoxicated group, BUN, AST and ALT levels, and histological criteria, such as infiltration of inflammatory cells, accumulation of reticulocytes, nuclear pyknosis of hepatocyte and sinusoidal dilation, significantly increased. In this group, ALB secretion significantly decreased, while AFP expression significantly increased. Both UCSCs and HLCs encapsulated in alginate scaffolds effectively attenuated biochemical tests, improved liver cytoarchitecture, increased expression of ALB and reduced AFP expression.
Conclusion: Finding of the present study indicated that encapsulation of UCSCs or HLCs in alginate mannuronic scaffolds effectively improve CCl4-induced ALF.


Main Subjects

1. Sun K, Xie X, Xie J, Jiao S, Chen X, Zhao X, et al. Cell-based therapy for acute and chronic liver failures: distinct diseases, different choices. Sci Rep 2014; 4:1-9.
2. Bernal W, Auzinger G, Dhawan A, Wendon J. Acute liver failure. Lancet 2010; 376:190-201.
3. Russo FP, Parola M. Stem cells in liver failure. Best Pract Res Clin Gastroenterol 2012; 26:35-45.
4. Sipe JD. Tissue engineering and reparative medicine. Ann N Y Acad of Sci 2002; 961:1-9.
5. Zhao Q, Ren H, Zhu D, Han Z. Stem/progenitor cells in liver injury repair and regeneration. Biol Cell 2009; 101:557-571.
6. Fu YS, Cheng YC, Lin MYA, Cheng H, Chu PM, Chou SC, et al. Conversion of human umbilical cord mesenchymal stem cells in Wharton’s jelly to dopaminergic neurons in vitro: potential therapeutic application for Parkinsonism. Stem cells 2006; 24:115-124.
7. Wu XB, Tao R. Hepatocyte differentiation of mesenchymal stem cells. Hepatobiliary Pancreat Dis Int 2012; 11:360-371.
8. Li P, Zhang J, Liu J, Ma H, Liu J, Lie P, et al. Promoting the recovery of injured liver with poly (3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) scaffolds loaded with umbilical cord-derived mesenchymal stem cells. Tissue Eng Part A 2014; 21:603-615.
9. Aleahmad F, Ebrahimi S, Salmannezhad M, Azarnia M, Jaberipour M, Hoseini M, et al. Heparin/collagen 3D scaffold accelerates hepatocyte differentiation of Wharton’s jelly-derived mesenchymal stem cells. Tissue Eng Regen Med 2017; 14:443-452.
10. Fang S, Qiu Yd, Mao L, Shi Xl, Yu DC, Ding Yt. Differentiation of embryoid-body cells derived from embryonic stem cells into hepatocytes in alginate microbeads in vitro. Acta Pharmacol Sin 2007; 28:1924-1930.   
11. Gautier A, Carpentier B, Dufresne M, Vu Dinh Q, Paullier P, Legallais C. Impact of alginate type and bead diameter on mass transfers and the metabolic activities of encapsulated C3A cells in bioartificial liver applications. Eur Cell Mater 2011; 21:94-106.
12. Ka    halil M, Shariat Panahi A, Tootle R, Ryder T, Mc Closkey P, Roberts E, et al. Human hepatocyte cell lines proliferating as cohesive spheroid colonies in alginate markedly upregulate both synthetic and detoxificatory liver function. J Hepatolo 2001; 34:68-77.
13. Kuhlmann WD, Peschke P. Hepatic progenitor cells, stem cells, and AFP expression in models of liver injury. Int J Exp Pathol 2006; 87:343-359.
14. Aoki T, Jin Z, Nishino N, Kato H, Shimizu Y, Niiya T, et al. Intrasplenic transplantation of encapsulated hepatocytes decreases mortality and improves liver functions in fulminant hepatic failure from 90% partial hepatectomy in rats. Transplantation 2005; 79:783-790.
15. Hashemitabar M, Allahbakhshi E, Tabande MR, Orazizadeh M, Dehbashi FN, Azandeh S, et al. Isolation and characterization of human umbilical cord mesenchymal stem cells and their differentiation into Pdx-1+ Cells. Int J Biomed Sci 2015; 8:780.
16. Chen MJ, Lu Y, Simpson NE, Beveridge MJ, Elshikha AS, Akbar MA, et al. In situ transplantation of alginate bioencapsulated adipose tissues derived stem cells (ADSCs) via hepatic injection in a mouse model. PLos One 2015; 10:e0138184.
17. Yoon HH, Jung BY, Seo YK, Song KY, Park JK. In vitro hepatic differentiation of umbilical cord-derived mesenchymal stem cell. Process Biochem 2010; 45:1857-1864.
18. Soleimani M, Khorsandi L, Atashi A, Nejaddehbashi F. Chondrogenic differentiation of human umbilical cord blood-derived unrestricted somatic stem cells on A 3D beta-tricalcium phosphate-alginate-gelatin scaffold. Cell J 2014; 16:43-52.
19. Saif A, Sarhan O, Mohamed Elmogy M, Mutwally H. Hepatoprotective effects of Zamzam water against Carbon Tetrachloride induced liver damage in Rats: Biochemical, Histopathological and molecular evidences. Life Sci 2014; 11:300-308.
20. Suvarna KS, Layton C, Bancroft JD. Bancroft’s Theory and Practice of Histological Techniques E-Book: Elsevier Health Sciences; 2018.
21. Ishak K, Baptista A, Bianchi L, Callea F, De Groote J, Gudat F, et al. Histological grading and staging of chronic hepatitis. J hepatol 1995; 22:696-699.
22. Azandeh S, Gharravi AM, Orazizadeh M, Khodadi A, Hashemi Tabar M. Improvement of mesenchymal stem cell differentiation into the endoderm lineage by four step sequential method in biocompatible biomaterial. Bioimpacts 2016; 6:9-13.
23. Hu C, Li L.  In vitro and in vivo hepatic differentiation of adult somatic stem cells and extraembryonic stem cells for treating end stage liver diseases. Stem Cells Int 2015; 871-972.
24. Seo MJ, Suh SY, Bae YC, Jung JS. Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun 2005; 32:258-264.
25. Gowda S, Desai PB, Hull VV, Math AAK, Vernekar SN, Kulkarni SS. A review on laboratory liver function tests. Pan Afr Med J 2009; 3:1-11.
26. Lin JS, Zhou L, Sagayaraj A, Jumat NHB, Choolani M, Chan JKY, et al. Hepatic differentiation of human amniotic epithelial cells and in vivo therapeutic effect on animal model of cirrhosis. J Gastroenterol Hepatol 2015; 30:1673-1682.
27. Li J, Zhang L, Xin J, Jiang L, Zhang T, Jin L, et al. Immediate intraportal transplantation of human bone marrow mesenchymal stem cells prevents death from fulminant hepatic failure in pigs. Hepatology 2012;  56:1044-1052.
28. Aldridge V, Garg A, Davies N, Bartlett DC, Youster J, Beard H, et al. Human mesenchymal stem cells are recruited to injured liver in a β1‐integrin and CD44 dependent manner. Hepatology 2012; 56:1063-1073.
29. Zhou R, Li Z, He C, Li R, Xia H, Li C, et al. Human umbilical cord mesenchymal stem cells and derived hepatocyte-like cells exhibit similar therapeutic effects on an acute liver failure mouse model. PLos One 2014; 9:e104392.
30. Ramanathan R, Pettinato G, Beeston JT, Lee DD, WenX, Mangino MJ, et al. Transplantation of human stem cell-derived hepatocytes in an animal model of acute liver failure. Surgery 2015; 158:349-359.
31. Asgari S, Moslem M, Bagheri-Lankarani K, Pournasr B, Miryounesi M, Baharvand H. Differentiation and transplantation of human induced pluripotent stem cell-derived hepatocyte-like cells. Stem Cell Rev 2013; 9:493-504.
32. Elmahdy NA, Sokar SS, Salem ML, Sarhan NI, Abou Elela SH. Anti-fibrotic potential of human umbilical cord mononuclear cells and mouse bone marrow cells in CCl4-induced liver fibrosis in mice. Biomed Pharmacother 2017; 89:1378-1386.
33. Deng L, Liu G, Wu X, Wang Y, Tong M, Liu B, et al. Adipose derived mesenchymal stem cells efficiently rescue carbon tetrachloride-induced acute liver failure in mouse. Sci World J 2014; 2014:1-8.
34. Cienfuegos J, Rotellar F, Baixauli J, Martínez Regueira F, Pardo F, Hernández Lizoáin JL. Liver regeneration the best kept secret, A model of tissue injury response. Rev Esp Enferm Dig 2014; 106:171-194.
35. Khodabandeh Z, Vojdani Z, Talaei-Khozani T, Jaberipour M, Hosseini A, Bahmanpour S. Comparison of the expression of hepatic genes by human wharton’s jelly mesenchymal stem cells cultured in 2D and 3D collagen culture systems. Iran J Med Sci 2016; 41:28-36.
36. Burra P, Arcidiacono D, Bizzaro D, Chioato T, Di Liddo R, Banerjee A, et al. Systemic administration of a novel human umbilical cord mesenchymal stem cells population accelerates the resolution of acute liver injury. BMC gastroenterol 2012; 12:1-16.
37. Tuo L, Zeng W, Xue H, Wu, X. Umbilical cord mesenchymal stem cells and their association with liver fibrosis. Zhonghua Gan Zang Bing Za Zhi 2017; 25:65-68.
38. Liu Z, Meng F, Li C, Zhou X, Zeng  X, He Y, et al. Human umbilical cord mesenchymal stromal cells rescue mice from acetaminophen-induced acute liver failure. Cytotherapy 2014; 16:1207-1219.
39. Umehara Y, Hakamada K, Seino K, Aoki K, Toyoki Y, Sasaki, M. Improved survival and ammonia metabolism by intraperitoneal transplantation of microencapsulated hepatocytes in totally hepatectomized rats. Surgery 2001;130:513-520.
40. Chen YX, Zeng ZC, Sun J, Zeng HY, Huang Y, Zhang ZY. Mesenchymal stem cell–conditioned medium prevents radiation-induced liver injury by inhibiting inflammation and protecting sinusoidal endothelial cells. J Radiat Res 2015;56:700-708.