title: Culturing on decellularized extracellular matrix enhances antioxidant properties of human umbilical cord-derived mesenchymal stem cells.
abstract: Human umbilical cord-derived mesenchymal stem cells (UC-MSCs) have attracted great interest in clinical application because of their regenerative potential and their lack of ethical issues. Our previous studies showed that decellularized cell-deposited extracellular matrix (ECM) provided an in vivo-mimicking microenvironment for MSCs and facilitated in vitro cell expansion. This study was conducted to analyze the cellular response of UC-MSCs when culturing on the ECM, including reactive oxygen species (ROS), intracellular antioxidative enzymes, and the resistance to exogenous oxidative stress. After decellularization, the architecture of cell-deposited ECM was characterized as nanofibrous, collagen fibrils and the matrix components were identified as type I and III collagens, fibronectin, and laminin. Compared to tissue culture polystyrene (TCPS) plates, culturing on ECM yielded a 2-fold increase of UC-MSC proliferation and improved the percentage of cells in the S phase by 2.4-fold. The levels of intracellular ROS and hydrogen peroxide (H2O2) in ECM-cultured cells were reduced by 41.7% and 82.9%, respectively. More importantly, ECM-cultured UC-MSCs showed enhanced expression and activity of intracellular antioxidative enzymes such as superoxide dismutase and catalase, up-regulated expression of silent information regulator type 1, and suppressed phosphorylation of p38 mitogen-activated protein kinase. Furthermore, a continuous treatment with exogenous 100μM H2O2 dramatically inhibited osteogenic differentiation of UC-MSCs cultured on TCPS, but culturing on ECM retained the differentiation capacity for matrix mineralization and osteoblast-specific marker gene expression. Collectively, by providing sufficient cell amounts and enhancing antioxidant capacity, decellularized ECM can be a promising cell culture platform for in vitro expansion of UC-MSCs.
authors.affiliation: School of Engineering, Sun Yat-sen University, Guangzhou 510006, China.
authors.forename: Xiaozhen
authors.initials: X
authors.lastname: Liu
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Long
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Pan
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Wenguo
authors.initials: W
authors.lastname: Cui
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Mao
authors.initials: M
authors.lastname: Li
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Zong-Ping
authors.initials: ZP
authors.lastname: Luo
authors.affiliation: Stem Cell and Tissue Engineering Laboratory, Department of Orthopaedics, West Virginia University, Morgantown, WV 26506, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Pei
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Huilin
authors.initials: H
authors.lastname: Yang
authors.affiliation: School of Engineering, Sun Yat-sen University, Guangzhou 510006, China. Electronic address: gongyih@mail.sysu.edu.cn.
authors.forename: Yihong
authors.initials: Y
authors.lastname: Gong
authors.affiliation: Orthopaedic Institute, Soochow University, Suzhou 215007, China; Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou 215006, China. Electronic address: fanhe@suda.edu.cn.
authors.forename: Fan
authors.initials: F
authors.lastname: He
mesh_terms: D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism*; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D005109:Extracellular Matrix / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D010012:Osteogenesis; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D016196:S Phase; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology
chemical_list: D000975:Antioxidants; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase
keywords: Antioxidative enzymes; Extracellular matrix; Mesenchymal stem cells; Osteogenesis; Reactive oxygen species
references.citation: Mol Cell. 2007 Apr 13;26(1):1-14
references.pmid: 17434122
references.citation: Free Radic Biol Med. 2014 Mar;68:234-46
references.pmid: 24374373
references.citation: Stem Cells Dev. 2010 Dec;19(12):1895-910
references.pmid: 20367285
references.citation: Matrix Biol. 2009 Jan;28(1):44-52
references.pmid: 19049867
references.citation: Spine (Phila Pa 1976). 2012 Mar 15;37(6):459-69
references.pmid: 21540772
references.citation: J Cell Biochem. 2003 Mar 1;88(4):783-93
references.pmid: 12577312
references.citation: Cell Biol Toxicol. 2006 Jan;22(1):39-46
references.pmid: 16463018
references.citation: Free Radic Biol Med. 2002 Aug 1;33(3):337-49
references.pmid: 12126755
references.citation: Rheumatology (Oxford). 2005 Jan;44(1):7-16
references.pmid: 15292527
references.citation: J Biomed Sci. 2010 Jul 12;17:56
references.pmid: 20624296
references.citation: Biomaterials. 2012 Jun;33(18):4480-9
references.pmid: 22459197
references.citation: Antioxid Redox Signal. 2013 May 20;18(15):1895-905
references.pmid: 23088254
references.citation: Placenta. 1997 Jan;18(1):53-64
references.pmid: 9032810
references.citation: Tissue Eng Part A. 2009 Dec;15(12):3809-21
references.pmid: 19545204
references.citation: Biochem Biophys Res Commun. 2013 Aug 23;438(2):270-6
references.pmid: 23891692
references.citation: Redox Rep. 2004;9(1):37-48
references.pmid: 15035826
references.citation: Arthritis Rheum. 2013 Feb;65(2):378-87
references.pmid: 23138846
references.citation: Antioxid Redox Signal. 2014 Apr 20;20(12):1881-90
references.pmid: 23066813
references.citation: Blood. 2010 Jan 21;115(3):443-52
references.pmid: 19797522
references.citation: Curr Med Chem. 2003 Oct;10(20):2123-45
references.pmid: 12871089
references.citation: PLoS One. 2011;6(10):e25990
references.pmid: 21998741
references.citation: Biochem Biophys Res Commun. 2008 Jul 18;372(1):51-6
references.pmid: 18485895
references.citation: PLoS One. 2008 May 21;3(5):e2213
references.pmid: 18493317
references.citation: Stem Cells. 2008 Jul;26(7):1749-57
references.pmid: 18467662
references.citation: Stem Cells Dev. 2013 Mar 15;22(6):889-900
references.pmid: 23092115
references.citation: Free Radic Biol Med. 2001 Mar 15;30(6):686-98
references.pmid: 11295367
references.citation: Mater Sci Eng C Mater Biol Appl. 2014 Jul 1;40:316-23
references.pmid: 24857499
references.citation: Tissue Eng Part A. 2011 Dec;17(23-24):3067-76
references.pmid: 21740327
references.citation: World J Stem Cells. 2014 Apr 26;6(2):195-202
references.pmid: 24772246
references.citation: Nat Rev Rheumatol. 2013 Oct;9(10):584-94
references.pmid: 23881068
references.citation: Curr Stem Cell Res Ther. 2013 Jan;8(1):39-45
references.pmid: 23278911
references.citation: J Pineal Res. 2015 Sep;59(2):190-205
references.pmid: 25975679
references.citation: Science. 1999 Apr 2;284(5411):143-7
references.pmid: 10102814
references.citation: Stem Cells Dev. 2010 Jul;19(7):1095-107
references.pmid: 19737070
references.citation: J Pineal Res. 2013 Apr;54(3):303-12
references.pmid: 23110400
references.citation: J Cell Physiol. 2012 May;227(5):2163-74
references.pmid: 21792932
references.citation: Nat Rev Rheumatol. 2015 Jan;11(1):35-44
references.pmid: 25266449
references.citation: Biomed Res Int. 2013;2013:406871
references.pmid: 23991414
references.citation: J Biol Chem. 2008 May 30;283(22):15319-27
references.pmid: 18378684
references.citation: Life Sci. 2014 Jun 6;105(1-2):1-6
references.pmid: 24657895
references.citation: J Pineal Res. 2013 Aug;55(1):14-25
references.pmid: 23488678
references.citation: Stem Cells. 2006 May;24(5):1226-35
references.pmid: 16424399
references.citation: J Mol Med (Berl). 2012 Apr;90(4):389-400
references.pmid: 22038097
references.citation: Int J Nanomedicine. 2012;7:5091-106
references.pmid: 23049256
references.citation: Biomaterials. 2014 Jan;35(2):642-53
references.pmid: 24148243
references.citation: Adv Exp Med Biol. 2010;694:126-37
references.pmid: 20886761
references.citation: Stem Cells. 2008 Mar;26(3):591-9
references.pmid: 18065397
references.citation: Mech Ageing Dev. 2002 Nov;123(12):1597-604
references.pmid: 12470897
references.citation: Oxid Med Cell Longev. 2013;2013:474931
references.pmid: 24062878
references.citation: J Endocrinol. 2014 Nov;223(2):167-80
references.pmid: 25210047
references.citation: Regen Med. 2009 May;4(3):423-33
references.pmid: 19438317