Adipose tissue-derived mesenchymal stem cells differentiated into hepatocyte-like cells in vivo and in vitro

Yin,Libo; Zhu,Yuhua; Yang,Jiangang; Ni,Yijiang; Zhou,Zhao; Chen,Yu; Wen,Lixing

doi:10.3892/mmr.2014.2935

Adipose tissue-derived mesenchymal stem cells differentiated into hepatocyte-like cells in vivo and in vitro

Authors:
- Libo Yin
- Yuhua Zhu
- Jiangang Yang
- Yijiang Ni
- Zhao Zhou
- Yu Chen
- Lixing Wen
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Affiliations: Department of Traumatic Surgery, Changzhou No. 2 People's Hospital, Nanjing Medical University, Changzhou, Jiangsu 213000, P.R. China
Published online on: November 13, 2014 https://doi.org/10.3892/mmr.2014.2935
Pages: 1722-1732
Copyright: © Yin et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Cell‑based therapy is a potential alternative to liver transplantation. The goal of the present study was to examine the in vivo and in vitro hepatic differentiation potential of adipose tissue‑derived mesenchymal stem cells (AT‑MSCs) and to explore its therapeutic use. AT‑MSCs were isolated and cultured with hepatic differentiation medium. Bioactivity assays were used to study the properties of AT‑MSCs. The morphology of differentiated AT‑MSCs in serum‑free hepatic differentiation medium changed into polygonal epithelial cells, while the morphology of AT‑MSCs in a similar medium containing 2% fetal bovine serum remained unchanged. The differentiated cells cultured without serum showed hepatocyte‑like cell morphology and hepatocyte‑specific markers, including albumin (ALB) and α‑fetoprotein. The bioactivity assays revealed that hepatocyte‑like cells could take up low‑density lipoprotein (LDL) and store glycogen. Furthermore, trichostatin A (TSA) enhanced ALB production and LDL uptake by the hepatocyte‑like cells, analogous to the functions of human liver cells. ALB was detected in the livers of the CCl4‑injured mice one month post‑transplantation. This suggested that transplantation of the human AT‑MSCs could relieve the impairment of acute CCl4‑injured livers in nude mice. This therefore implied that adipose tissue was a source of multipotent stem cells which had the potential to differentiate into mature, transplantable hepatocyte‑like cells in vivo and in vitro. In addition, the present study determined that TSA was essential to promoting differentiation of human MSC towards functional hepatocyte‑like cells. The relief of liver injury following treatment with AT‑MSCs suggested their potential as a novel therapeutic method for liver disorders or injury.

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1	van Poll D, Parekkadan B, Cho CH, et al: Mesenchymal stem cell-derived molecules directly modulate hepatocellular death and regeneration in vitro and in vivo. Hepatology. 47:1634–1643. 2008. View Article : Google Scholar : PubMed/NCBI
2	Kashofer K and Bonnet D: Gene therapy progress and prospects: stem cell plasticity. Gene Ther. 12:1229–1234. 2005. View Article : Google Scholar : PubMed/NCBI
3	Kern S, Eichler H, Stoeve J, Klüter H and Bieback K: Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 24:1294–1301. 2006. View Article : Google Scholar : PubMed/NCBI
4	Devine SM, Cobbs C, Jennings M, Bartholomew A and Hoffman R: Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood. 101:2999–3001. 2003. View Article : Google Scholar
5	Tepliashin AS, Chupikova NI, Korzhikova SV, et al: Comparative analysis of cell populations with a phenotype similar to that of mesenchymal stem cells derived from subcutaneous fat. Tsitologiia. 47:637–643. 2005.(in Russian).
6	Malhi H, Irani AN, Gagandeep S and Gupta S: Isolation of human progenitor liver epithelial cells with extensive replication capacity and differentiation into mature hepatocytes. J Cell Sci. 115:2679–2688. 2002.PubMed/NCBI
7	Young HE, Steele TA, Bray RA, et al: Human reserve pluripotent mesenchymal stem cells are present in the connective tissues of skeletal muscle and dermis derived from fetal, adult, and geriatric donors. Anat Rec. 264:51–62. 2001. View Article : Google Scholar : PubMed/NCBI
8	Lowes KN, Croager EJ, Olynyk JK, Abraham LJ and Yeoh GC: Oval cell-mediated liver regeneration: Role of cytokines and growth factors. J Gastroenterol Hepatol. 18:4–12. 2003. View Article : Google Scholar : PubMed/NCBI
9	Zuk PA, Zhu M, Ashjian P, et al: Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 13:4279–4295. 2002. View Article : Google Scholar : PubMed/NCBI
10	Lagasse E, Connors H, Al-Dhalimy M, et al: Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med. 6:1229–1234. 2000. View Article : Google Scholar : PubMed/NCBI
11	Alison MR, Poulsom R, Jeffery R, et al: Hepatocytes from non-hepatic adult stem cells. Nature. 406:2572000. View Article : Google Scholar : PubMed/NCBI
12	Herzog EL, Chai L and Krause DS: Plasticity of marrow-derived stem cells. Blood. 102:3483–3493. 2003. View Article : Google Scholar : PubMed/NCBI
13	Vassilopoulos G, Wang PR and Russell DW: Transplanted bone marrow regenerates liver by cell fusion. Nature. 422:901–904. 2003. View Article : Google Scholar : PubMed/NCBI
14	Wang X, Willenbring H, Akkari Y, et al: Cell fusion is the principal source of bone-marrow-derived hepatocytes. Nature. 422:897–901. 2003. View Article : Google Scholar : PubMed/NCBI
15	Wagner W, Wein F, Seckinger A, et al: Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp Hematol. 33:1402–1416. 2005. View Article : Google Scholar : PubMed/NCBI
16	Snykers S, Vanhaecke T, De Becker A, et al: Chromatin remodeling agent trichostatin A: a key-factor in the hepatic differentiation of human mesenchymal stem cells derived of adult bone marrow. BMC Dev Biol. 7:242007. View Article : Google Scholar : PubMed/NCBI
17	Yamamoto H, Quinn G, Asari A, et al: Differentiation of embryonic stem cells into hepatocytes: biological functions and therapeutic application. Hepatology. 37:983–993. 2003. View Article : Google Scholar : PubMed/NCBI
18	Kim JW, Kim SY, Park SY, et al: Mesenchymal progenitor cells in the human umbilical cord. Ann Hematol. 83:733–738. 2004. View Article : Google Scholar : PubMed/NCBI
19	Reyes M, Lund T, Lenvik T, et al: Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood. 98:2615–2625. 2001. View Article : Google Scholar : PubMed/NCBI
20	Teratani T, Yamamoto H, Aoyagi K, et al: Direct hepatic fate specification from mouse embryonic stem cells. Hepatology. 41:836–846. 2005. View Article : Google Scholar : PubMed/NCBI
21	Yamamoto Y, Teratani T, Yamamoto H, et al: Recapitulation of in vivo gene expression during hepatic differentiation from murine embryonic stem cells. Hepatology. 42:558–567. 2005. View Article : Google Scholar : PubMed/NCBI
22	Baertschiger RM, Serre-Beinier V, Morel P, et al: Fibrogenic potential of human multipotent mesenchymal stromal cells in injured liver. PLoS One. 4:e66572009. View Article : Google Scholar : PubMed/NCBI
23	Chien CC, Yen BL, Lee FK, et al: In vitro differentiation of human placenta-derived multipotent cells into hepatocyte-like cells. Stem Cells. 24:1759–1768. 2006. View Article : Google Scholar : PubMed/NCBI
24	Herold C, Ganslmayer M, Ocker M, et al: The histone-deacetylase inhibitor Trichostatin A blocks proliferation and triggers apoptotic programs in hepatoma cells. J Hepatol. 36:233–240. 2002. View Article : Google Scholar : PubMed/NCBI
25	Vinken M, Henkens T, Vanhaecke T, et al: Trichostatin a enhances gap junctional intercellular communication in primary cultures of adult rat hepatocytes. Toxicol Sci. 91:484–492. 2006. View Article : Google Scholar : PubMed/NCBI
26	Henkens T, Papeleu P, Elaut G, et al: Trichostatin A, a critical factor in maintaining the functional differentiation of primary cultured rat hepatocytes. Toxicol Appl Pharmacol. 218:64–71. 2007. View Article : Google Scholar
27	Bochev I, Elmadjian G, Kyurkchiev D, et al: Mesenchymal stem cells from human bone marrow or adipose tissue differently modulate mitogen-stimulated B-cell immunoglobulin production in vitro. Cell Biol Int. 32:384–393. 2008. View Article : Google Scholar : PubMed/NCBI
28	McIntosh K, Zvonic S, Garrett S, et al: The immunogenicity of human adipose-derived cells: temporal changes in vitro. Stem Cells. 24:1246–1253. 2006. View Article : Google Scholar : PubMed/NCBI
29	Battiwalla M and Hematti P: Mesenchymal stem cells in hematopoietic stem cell transplantation. Cytotherapy. 11:503–515. 2009. View Article : Google Scholar : PubMed/NCBI
30	Dinarello CA: The interleukin-1 family: 10 years of discovery. FASEB J. 8:1314–1325. 1994.PubMed/NCBI
31	Ortiz LA, Dutreil M, Fattman C, et al: Interleukin 1 receptor antagonist mediates the anti-inflammatory and antifibrotic effect of mesenchymal stem cells during lung injury. Proc Natl Acad Sci USA. 104:11002–11007. 2007. View Article : Google Scholar
32	Ponte AL, Marais E, Gallay N, et al: The in vitro migration capacity of human bone marrow mesenchymal stem cells: comparison of chemokine and growth factor chemotactic activities. Stem Cells. 25:1737–1745. 2007. View Article : Google Scholar : PubMed/NCBI
33	Potian JA, Aviv H, Ponzio NM, Harrison JS and Rameshwar P: Veto-like activity of mesenchymal stem cells: functional discrimination between cellular responses to alloantigens and recall antigens. J Immunol. 171:3426–3434. 2003. View Article : Google Scholar : PubMed/NCBI
34	Tang J, Wang J, Yang J, et al: Mesenchymal stem cells over-expressing SDF-1 promote angiogenesis and improve heart function in experimental myocardial infarction in rats. Eur J Cardiothorac Surg. 36:644–650. 2009. View Article : Google Scholar : PubMed/NCBI
35	Greco SJ and Rameshwar P: Microenvironmental considerations in the application of human mesenchymal stem cells in regenerative therapies. Biologics. 2:699–705. 2008.
36	Mouiseddine M, François S, Semont A, et al: Human mesenchymal stem cells home specifically to radiation-injured tissues in a non-obese diabetes/severe combined immunodeficiency mouse model. Br J Radiol. 80(Spec 1): S49–55. 2007. View Article : Google Scholar : PubMed/NCBI
37	Oyagi S, Hirose M, Kojima M, et al: Therapeutic effect of transplanting HGF-treated bone marrow mesenchymal cells into CCl₄-injured rats. J Hepatol. 44:742–748. 2006. View Article : Google Scholar : PubMed/NCBI
38	Pan MX, Hou WL, Zhang QJ, et al: Infusion of autologous mesenchymal stem cells prolongs the survival of dogs receiving living donor liver transplantation. J South Med Univ. (in Chinese).
39	Kuo YR, Goto S, Shih HS, et al: Mesenchymal stem cells prolong composite tissue allotransplant survival in a swine model. Transplantation. 87:1769–1777. 2009. View Article : Google Scholar : PubMed/NCBI
40	Matsuda Y, Matsumoto K, Ichida T and Nakamura T: Hepatocyte growth factor suppresses the onset of liver cirrhosis and abrogates lethal hepatic dysfunction in rats. J Biochem. 118:643–649. 1995.PubMed/NCBI
41	Silva GV, Litovsky S, Assad JA, et al: Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model. Circulation. 111:150–156. 2005. View Article : Google Scholar : PubMed/NCBI
42	Phinney DG and Prockop DJ: Concise review: mesenchymal stem/multipotent stromal cells: the state of transdifferentiation and modes of tissue repair - current views. Stem Cells. 25:2896–2902. 2007. View Article : Google Scholar : PubMed/NCBI
43	Krampera M, Cosmi L, Angeli R, et al: Role for interferon-gamma in the immunomodulatory activity of human bone marrow mesenchymal stem cells. Stem Cells. 24:386–398. 2006. View Article : Google Scholar
44	Rasmusson I, Uhlin M, Le Blanc K and Levitsky V: Mesenchymal stem cells fail to trigger effector functions of cytotoxic T lymphocytes. J Leukoc Biol. 82:887–893. 2007. View Article : Google Scholar : PubMed/NCBI