Restricted accessResearch articleFirst published online 2013-04
Effect of Antioxidant Supplementation on the Total Yield,Oxidative Stress Levels,and Multipotency of Bone Marrow-Derived Human Mesenchymal Stromal Cells
Bone marrow-derived multipotent mesenchymal stromal cells (MSCs) are the most frequently investigated cell type for potential regenerative strategies because they are relatively easy to isolate and are able to differentiate into several mesenchymal lineages. Unfortunately, during ex vivo culture, MSCs present gradual loss of differentiation potential and reduced clinical efficacy. Reactive oxygen species (ROS) are associated with oxidative damage and accumulate during MSC expansion. Because ROS are believed to be involved in the loss of multipotency, we hypothesized that compounds with antioxidant activity have the capacity to scavenge ROS, prevent cellular damage, and rescue culture-induced loss of multipotency. In this manuscript, we show that antioxidant supplementation can partially rescue the loss of alkaline phosphatase expression induced by oxidizing agents and increases the yield of hMSCs, when supplemented to a fresh bone marrow aspirate. Concomitantly, oxidative DNA damage and ROS levels in hMSCs were reduced by antioxidants. We conclude that antioxidant supplementation during MSC expansion reduces the DNA damage load and increases the MSC yield.
Get full access to this article
View all access options for this article.
References
1.
KadiyalaS.Culture-expanded, bone marrow-derived mesenchymal stem cells can regenerate a critical-sized segmental bone defect. Tissue Eng, 3:173. 1997.
2.
BruderS.P.et al.Mesenchymal stem cells in osteobiology and applied bone regeneration. Clin Orthop Relat Res355 Suppl:S247. 1998.
3.
Almeida-PoradaG.et al.Cotransplantation of human stromal cell progenitors into preimmune fetal sheep results in early appearance of human donor cells in circulation and boosts cell levels in bone marrow at later time points after transplantation. Blood, 95:3620. 2000.
4.
TseW.T.et al.Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation, 75:389. 2003.
5.
RingdenO.et al.Mesenchymal stem cells for treatment of therapy-resistant graft-versus-host disease. Transplantation, 81:1390. 2006.
6.
PittengerM.F.et al.Multilineage potential of adult human mesenchymal stem cells. Science, 284:143. 1999.
7.
PetiteH.et al.Tissue-engineered bone regeneration. Nat Biotechnol, 18:959. 2000.
8.
AgataH.et al.Characteristic change and loss of in vivo osteogenic abilities of human bone marrow stromal cells during passage. Tissue Eng Part A, 16:663. 2010.
9.
SiddappaR.et al.Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering. J Orthop Res, 25:1029. 2007.
10.
AlvesH.et al.A link between the accumulation of DNA damage and loss of multi-potency of human mesenchymal stromal cells. J Cell Mol Med, 14:2729. 2010.
11.
FaganJ.M., SleczkaB.G., SoharI.Quantitation of oxidative damage to tissue proteins. Int J Biochem Cell Biol, 31:751. 1999.
12.
KohenR., NyskaA.Oxidation of biological systems: oxidative stress phenomena, antioxidants, redox reactions, and methods for their quantification. Toxicol Pathol, 30:620. 2002.
13.
Dalle-DonneI.et al.Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta, 329:23. 2003.
14.
NikiE.et al.Lipid peroxidation: mechanisms, inhibition, and biological effects. Biochem Biophys Res Commun, 338:668. 2005.
15.
CadetJ.et al.Hydroxyl radicals and DNA base damage. Mutat Res, 424:9. 1999.
16.
FinkelT., HolbrookN.J.Oxidants, oxidative stress and the biology of ageing. Nature, 408:239. 2000.
17.
ValkoM.et al.Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol, 39:44. 2007.
18.
DudasS.P., ArkingR.A coordinate upregulation of antioxidant gene activities is associated with the delayed onset of senescence in a long-lived strain of Drosophila. J Gerontol A Biol Sci Med Sci, 50:B117. 1995.
19.
HarshmanL.G., HabererB.A.Oxidative stress resistance: a robust correlated response to selection in extended longevity lines of Drosophila melanogaster?J Gerontol A Biol Sci Med Sci, 55:B415. 2000.
20.
BalabanR.S., NemotoS., FinkelT.Mitochondria, oxidants, and aging. Cell, 120:483. 2005.
21.
KuH.H., BrunkU.T., SohalR.S.Relationship between mitochondrial superoxide and hydrogen peroxide production and longevity of mammalian species. Free Radic Biol Med, 15:621. 1993.
22.
MaggioD.et al.Marked decrease in plasma antioxidants in aged osteoporotic women: results of a cross-sectional study. J Clin Endocrinol Metab, 88:1523. 2003.
23.
BasuS.et al.Association between oxidative stress and bone mineral density. Biochem Biophys Res Commun, 288:275. 2001.
24.
de BoerJ.et al.Premature aging in mice deficient in DNA repair and transcription. Science, 296:1276. 2002.
25.
TynerS.D.et al.p53 mutant mice that display early ageing-associated phenotypes. Nature, 415:45. 2002.
26.
ItoK.et al.Regulation of oxidative stress by ATM is required for self-renewal of haematopoietic stem cells. Nature, 431:997. 2004.
27.
EbertR.et al.Selenium supplementation restores the antioxidative capacity and prevents cell damage in bone marrow stromal cells in vitro. Stem Cells, 24:1226. 2006.
28.
HockenberyD.M.et al.Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell, 75:241. 1993.
29.
JakubowskiW., BartoszG.2,7-dichlorofluorescin oxidation and reactive oxygen species: what does it measure?Cell Biol Int, 24:757. 2000.
30.
De BoerJ., WangH.J., Van BlitterswijkC.Effects of Wnt signaling on proliferation and differentiation of human mesenchymal stem cells. Tissue Eng, 10:393. 2004.
31.
BorsW., SaranM., MichelC.Pulse-radiolytic investigations of catechols and catecholamines. II. Reactions of Tiron with oxygen radical species. Biochim Biophys Acta, 582:537. 1979.
32.
ChingT.L., HaenenG.R., BastA.Cimetidine and other H2 receptor antagonists as powerful hydroxyl radical scavengers. Chem Biol Interact, 86:119. 1993.
33.
ValkoM.et al.Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact, 160:1. 2006.
34.
JoyeuxM.et al.tert-Butyl hydroperoxide-induced injury in isolated rat hepatocytes: a model for studying anti-hepatotoxic crude drugs. Planta Med, 56:171. 1990.
35.
FernandesE.R.et al.Hepatoprotective activity of xanthones and xanthonolignoids against tert-butylhydroperoxide-induced toxicity in isolated rat hepatocytes—comparison with silybin. Pharm Res, 12:1756. 1995.
36.
TsengT.H.et al.Protective effects of dried flower extracts of Hibiscus sabdariffa L. against oxidative stress in rat primary hepatocytes. Food Chem Toxicol, 35:1159. 1997.
37.
HalliwellB.Role of free radicals in the neurodegenerative diseases: therapeutic implications for antioxidant treatment. Drugs Aging, 18:685. 2001.
38.
NordbergJ., ArnerE.S.Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free Radic Biol Med, 31:1287. 2001.
QiW.et al.Chromium(III)-induced 8-hydroxydeoxyguanosine in DNA and its reduction by antioxidants: comparative effects of melatonin, ascorbate, and vitamin E. Environ Health Perspect, 108:399. 2000.
41.
LentonK.J.et al.Vitamin C augments lymphocyte glutathione in subjects with ascorbate deficiency. Am J Clin Nutr, 77:189. 2003.
42.
CarrA., FreiB.Does vitamin C act as a pro-oxidant under physiological conditions?FASEB J, 13:1007. 1999.
JiA.R.et al.Reactive oxygen species enhance differentiation of human embryonic stem cells into mesendodermal lineage. Exp Mol Med, 42:175. 2010.
45.
HiguchiM., DustingG.J., PeshavariyaH., JiangF., HsiaoS.T., ChanE., LiuG.S.Differentiation of human adipose-derived stem cells into fat involves reactive oxygen species and forkhead box O1 mediated upregulation of antioxidant enzymes. Stem Cells Dev, 2012[Epub ahead of print]10.1089/scd.2012.0306.
46.
SmithJ.et al.Redox state is a central modulator of the balance between self-renewal and differentiation in a dividing glial precursor cell. Proc Natl Acad Sci U S A, 97:10032. 2000.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
