This study investigated whether xenotransplantation of human Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) reduces thioacetamide (TAA)-induced mouse liver fibrosis and the underlying molecular mechanism.
Recipient NOD/SCID mice were injected intraperitoneally with TAA twice weekly for 6 weeks before initial administration of WJ-MSCs. Expression of regenerative and pro-fibrogenic markers in mouse fibrotic livers were monitored post cytotherapy. A hepatic stallate cell line HSC-T6 and isolated WJ-MSCs were used for in vitro adhesion, migration and mechanistic studies.
WJ-MSCs were isolated from human umbilical cords by an explant method and characterized by flow cytometry. A single infusion of WJ-MSCs to TAA-treated mice significantly reduced collagen deposition and ameliorated liver fibrosis after 2-week therapy. In addition to enhanced expression of hepatic regenerative factor, hepatocyte growth factor, and PCNA proliferative marker, WJ-MSC therapy significantly blunted pro-fibrogenic signals, including Smad2, RhoA, ERK. Intriguingly, reduction of plasma fibronectin (pFN) in fibrotic livers was noted in MSC-treated mice. In vitro studies further demonstrated that suspending MSCs triggered pFN degradation, soluble pFN conversely retarded adhesion of suspending MSCs onto type I collagen-coated surface, whereas pFN coating enhanced WJ-MSC migration across mimicked wound bed. Moreover, pretreatment with soluble pFN and conditioned medium from MSCs with pFN strikingly attenuated the response of HSC-T6 cells to TGF-β1-stimulation in Smad2 phosphorylation and RhoA upregulation.
These findings suggest that cytotherapy using WJ-MSCs may modulate hepatic pFN deposition for a better regenerative niche in the fibrotic livers and may constitute a useful anti-fibrogenic intervention in chronic liver diseases.
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Bissell DM. Hepatic fibrosis as wound repair: a progress report. J Gastroenterol. 1998;33:295–302.
Tsukada S, Parsons CJ, Rippe RA. Mechanisms of liver fibrosis. Clin Chim Acta. 2006;364:33–60.
Friedman SL. Molecular mechanisms of hepatic fibrosis and principles of therapy. J Gastroenterol. 1997;32:424–30.
Qian J, Niu M, Zhai X, Zhou Q, Zhou Y. β-Catenin pathway is required for TGF-β inhibition of PPARgamma expression in cultured hepatic stellate cells. Pharmacol Res. 2012;66:219–25.
Deng X, Deng L, Wang P, Cheng C, Xu K. Post-translational modification of CREB-1 decreases collagen I expression by inhibiting the TGF-β1 signaling pathway in rat hepatic stellate cells. Mol Med Rep. 2016;14:5751–9.
Okita K, Yamanaka S. Induction of pluripotency by defined factors. Exp Cell Res. 2010;316:2565–70.
McElreavey KD, Irvine AI, Ennis KT, McLean WH. Isolation, culture and characterisation of fibroblast-like cells derived from the Wharton’s jelly portion of human umbilical cord. Biochem Soc Trans. 1991;19:29S.
Takechi K, Kuwabara Y, Mizuno M. Ultrastructural and immunohistochemical studies of Wharton’s jelly umbilical cord cells. Placenta. 1993;14:235–45.
Kobayashi K, Kubota T, Aso T. Study on myofibroblast differentiation in the stromal cells of Wharton’s jelly: expression and localization of alpha-smooth muscle actin. Early Hum Dev. 1998;51:223–33.
Wang HS, Hung SC, Peng ST, Huang CC, Wei HM, Guo YJ, et al. Mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord. Stem Cells. 2004;22:1330–7.
Fong CY, Chak LL, Biswas A, Tan JH, Gauthaman K, Chan WK, et al. Human Wharton’s jelly stem cells have unique transcriptome profiles compared to human embryonic stem cells and other mesenchymal stem cells. Stem Cell Rev Rep. 2011;7:1–16.
Mitchell KE, Weiss ML, Mitchell BM, Martin P, Davis D, Morales L, et al. Matrix cells from Wharton’s jelly form neurons and glia. Stem Cells. 2003;21:50–60.
Chao KC, Chao KF, Fu YS, Liu SH. Islet-like clusters derived from mesenchymal stem cells in Wharton’s Jelly of the human umbilical cord for transplantation to control type 1 diabetes. PLoS One. 2008;3:e1451.
Conconi MT, Burra P, Di Liddo R, Calore C, Turetta M, Bellini S, et al. CD105(+) cells from Wharton’s jelly show in vitro and in vivo myogenic differentiative potential. Int J Mol Med. 2006;18:1089–96.
Wu KH, Zhou B, Lu SH, Feng B, Yang SG, Du WT, et al. In vitro and in vivo differentiation of human umbilical cord derived stem cells into endothelial cells. J Cell Biochem. 2007;100:608–16.
Zhang YN, Lie PC, Wei X. Differentiation of mesenchymal stromal cells derived from umbilical cord Wharton’s jelly into hepatocyte-like cells. Cytotherapy. 2009;11:548–58.
Anzalone R, Lo Iacono M, Corrao S, Magno F, Loria T, Cappello F, et al. New emerging potentials for human Wharton’s jelly mesenchymal stem cells: immunological features and hepatocyte-like differentiative capacity. Stem Cells Dev. 2010;19:423–38.
Tsai PC, Fu TW, Chen YM, Ko TL, Chen TH, Shih YH, et al. The therapeutic potential of human umbilical mesenchymal stem cells from Wharton’s jelly in the treatment of rat liver fibrosis. Liver Transpl. 2009;15:484–95.
Lin SZ, Chang YJ, Liu JW, Chang LF, Sun LY, Li YS, et al. Transplantation of human Wharton’s Jelly-derived stem cells alleviates chemically induced liver fibrosis in rats. Cell Transplant. 2010;19:1451–63.
Sakaida I, Terai S, Yamamoto N, Aoyama K, Ishikawa T, Nishina H, et al. Transplantation of bone marrow cells reduces CCl4-induced liver fibrosis in mice. Hepatology. 2004;40:1304–11.
Higashiyama R, Inagaki Y, Hong YY, Kushida M, Nakao S, Niioka M, et al. Bone marrow-derived cells express matrix metalloproteinases and contribute to regression of liver fibrosis in mice. Hepatology. 2007;45:213–22.
Sun CK, Chen CH, Kao YH, Yuen CM, Sheu JJ, Lee FY, et al. Bone marrow cells reduce fibrogenesis and enhance regeneration in fibrotic rat liver. J Surg Res. 2011;169:e15–26.
Sáez-Lara MJ, Frecha C, Martín F, Abadía F, Toscano M, Gil A, et al. Transplantation of human CD34+ stem cells from umbilical cord blood to rats with thioacetamide-induced liver cirrhosis. Xenotransplantation. 2006;13:529–35.
Kim YH, Cho KA, Park M, Kim HS, Park JW, Woo SY, et al. Conditioned medium from tonsil-derived mesenchymal stem cells relieves CCl4-induced liver fibrosis in mice. Tissue Eng Regen Med. 2019;16:51–8.
De Bruyn C, Najar M, Raicevic G, Meuleman N, Pieters K, Stamatopoulos B, et al. A rapid, simple, and reproducible method for the isolation of mesenchymal stromal cells from Wharton’s jelly without enzymatic treatment. Stem Cells Dev. 2011;20:547–57.
Chang CY, Chen PH, Li CJ, Lu SC, Lin YC, Lee PH, et al. Isolation and characterization of mesenchymal stem cells derived from human umbilical cord blood mononuclear cells. E-Da Medical Journal. 2016;3:1–13.
Kao YH, Chen CL, Jawan B, Chung YH, Sun CK, Kuo SM, et al. Upregulation of hepatoma-derived growth factor is involved in murine hepatic fibrogenesis. J Hepatol. 2010;52:96–105.
Junqueira LC, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J. 1979;11:447–55.
Goodman ZD, Becker RL Jr, Pockros PJ, Afdhal NH. Progression of fibrosis in advanced chronic hepatitis C: evaluation by morphometric image analysis. Hepatology. 2007;45:886–94.
Chang YC, Kao YH, Hu DN, Tsai LY, Wu WC. All-trans retinoic acid remodels extracellular matrix and suppresses laminin-enhanced contractility of cultured human retinal pigment epithelial cells. Exp Eye Res. 2009;88:900–9.
Wu WC, Lai YH, Hsieh MC, Chang YC, Wu MH, Wu HJ, et al. Pleiotropic role of atorvastatin in regulation of human retinal pigment epithelial cell behaviors in vitro. Exp Eye Res. 2011;93:842–51.
Kao YH, Chen PH, Wu TY, Lin YC, Tsai MS, Lee PH, et al. Lipopolysaccharides induce Smad2 phosphorylation through PI3K/Akt and MAPK cascades in HSC-T6 hepatic stellate cells. Life Sci. 2017;184:37–46.
Jiang Y, Xu W, Lu J, He F, Yang X. Invasiveness of hepatocellular carcinoma cell lines: contribution of hepatocyte growth factor, c-met, and transcription factor Ets-1. Biochem Biophys Res Commun. 2001;286:1123–30.
Sun XE, Zhang XQ, Liu MM. Effect of bone marrow mesenchymal stem cells on the TGF-β1/Smad signaling pathway of hepatic stellate. Genet Mol Res. 2015;14:8744–54.
Ohara M, Ohnishi S, Hosono H, Yamamoto K, Yuyama K, Nakamura H, et al. Extracellular vesicles from amnion-derived mesenchymal stem cells ameliorate hepatic inflammation and fibrosis in rats. Stem Cells Int. 2018;2018:3212643.
Qin S, Jiang H, Su S, Wang D, Liang Z, Zhang J, et al. Inhibition of hepatic stellate cell proliferation by bone marrow mesenchymal stem cells via regulation of the cell cycle in rat. Exp Ther Med. 2012;4:375–80.
An SY, Jang YJ, Lim HJ, Han J, Lee J, Lee G, et al. Milk fat globule-EGF factor 8, secreted by mesenchymal stem cells, protects against liver fibrosis in mice. Gastroenterology. 2017;152:1174–86.
Huang B, Cheng X, Wang H, Huang W, la Ga HuZ, Wang D, et al. Mesenchymal stem cells and their secreted molecules predominantly ameliorate fulminant hepatic failure and chronic liver fibrosis in mice respectively. J Transl Med. 2016;14:45.
Qiao H, Zhou Y, Qin X, Cheng J, He Y, Jiang Y. NADPH oxidase signaling pathway mediates mesenchymal stem cell-induced inhibition of hepatic stellate cell activation. Stem Cells Int. 2018;2018:1239143.
Wang J, Sun M, Liu W, Li Y, Li M. Stem cell-based therapies for liver diseases: an overview and update. Tissue Eng Regen Med. 2019;16:107–18.
To WS, Midwood KS. Plasma and cellular fibronectin: distinct and independent functions during tissue repair. Fibrogenesis Tissue Repair. 2011;4:21.
Moretti FA, Chauhan AK, Iaconcig A, Porro F, Baralle FE, Muro AF. A major fraction of fibronectin present in the extracellular matrix of tissues is plasma-derived. J Biol Chem. 2007;282:28057–62.
Kwon AH, Inada Y, Uetsuji S, Yamamura M, Hioki K, Yamamoto M. Response of fibronectin to liver regeneration after hepatectomy. Hepatology. 1990;11:593–8.
Kwon AH, Uetsuji S, Yamamura M, Hioki K, Yamamoto M. Effect of administration of fibronectin or aprotinin on liver regeneration after experimental hepatectomy. Ann Surg. 1990;211:295–300.
Moriyama T, Aoyama H, Ohnishi S, Imawari M. Protective effects of fibronectin in galactosamine-induced liver failure in rats. Hepatology. 1986;6:1334–9.
Oh E, Pierschbacher M, Ruoslahti E. Deposition of plasma fibronectin in tissues. Proc Natl Acad Sci U S A. 1981;78:3218–21.
Pujades C, Forsberg E, Enrich C, Johansson S. Changes in cell surface expression of fibronectin and fibronectin receptor during liver regeneration. J Cell Sci. 1992;102:815–20.
Altrock E, Sens C, Wuerfel C, Vasel M, Kawelke N, Dooley S, et al. Inhibition of fibronectin deposition improves experimental liver fibrosis. J Hepatol. 2015;62:625–33.
De Luna-Saldivar MM, Marino-Martinez IA, Franco-Molina MA, Rivera-Morales LG, Alarcón-Galván G, Cordero-Pérez P, et al. Advantages of adipose tissue stem cells over CD34(+) mobilization to decrease hepatic fibrosis in Wistar rats. Ann Hepatol. 2019;18:620–6.
Cho IJ, Kim YW, Han CY, Kim EH, Anderson RA, Lee YS, et al. E-cadherin antagonizes transforming growth factor beta1 gene induction in hepatic stellate cells by inhibiting RhoA-dependent Smad3 phosphorylation. Hepatology. 2010;52:2053–64.
Eliazer S, Muncie JM, Christensen J, Sun X, D’Urso RS, Weaver VM, et al. Wnt4 from the niche controls the mechano-properties and quiescent state of muscle stem cells. Cell Stem Cell. 2019;25:654–65.e4.
Bruck R, Hershkoviz R, Lider O, Aeed H, Zaidel L, Matas Z, et al. Inhibition of experimentally-induced liver cirrhosis in rats by a nonpeptidic mimetic of the extracellular matrix-associated Arg–Gly–Asp epitope. J Hepatol. 1996;24:731–8.
Hayes-Jordan A, Wang YX, Walker P, Cox CS. Mesenchymal stromal cell dependent regression of pulmonary metastasis from Ewing’s. Front Pediatr. 2014;2:44.
Barker EA, Smuckler EA. Nonhepatic thioacetamide injury. II. The morphologic features of proximal renal tubular injury. Am J Pathol. 1974;74:575–90.
The authors would like to dedicate this article to late Professor Daniel Tzu-Bi Shih in memory of his earnestness and generosity in disseminating his knowledge and sharing his expertise with his fellow researchers. This study was supported by the Grants from E-Da Hospital (EDAHP102043 to CYC; NCKUEDA10513 to YHK; EDAHT107047 to CKS). YHK, CYC and CKS conceived the study, supervised the experiments and finalized the draft. YHK and YCL wrote the paper. YCL, CWL and PHC performed the experiments. PHL, TST, YCC, and MHC contributed to analysis tools and analyzed the data. All authors have reviewed the manuscript and approved submission.
Conflict of interest
The authors declare that they have no conflict of interest.
The study protocol was approved by the institutional reviewing board of E-Da Hospital (IRB No. EMRP26100N). Informed consent was confirmed by the IRB. The animal studies were performed after receiving approval of the Institutional Animal Care and Use Committee at E-Da Hospital (IACUC Approval No. IACUC-101017).
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Kao, Y., Lin, Y., Lee, P. et al. Infusion of Human Mesenchymal Stem Cells Improves Regenerative Niche in Thioacetamide-Injured Mouse Liver. Tissue Eng Regen Med (2020). https://doi.org/10.1007/s13770-020-00274-4
- Human umbilical cord
- Liver fibrosis
- Plasma fibronectin
- Wharton’s jelly tissue