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| The Effect of Reduced Glutathione to the Chondrogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells |
| LUO Er-mei, YU Li, ZHANG Jia-wen, LIU Jing |
| Jinan University, Guangzhou 510632, China |
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Abstract Objective: Reduced glutathione (GSH) were discussed whether GSH could promote the chondrogenic induction ability of human umbilical cord mesenchymal stem cells (hUC-MSCs). Methods: hUC-MSCs were isolated from human umbilical cord and their specificity was identified, then, induced into cartilage-like cells in chondrogenic induction medium with tansforming growth factor beta 1 (TGF-β1), especially with GSH. The morphological change before and after induction was observed through inverted phase contrast microscope, Type Ⅱcollagen(COL2A1) and glycosaminoglycan (GAG) were analyzed qualitatively by toluidine blue and immunofluorescence technique, respectively, the contents of COL2A1 and GAG were estimated from the determination of hydroxyproline content and Alcian Blue method separately. The mRNA expressions of GAG and COL2A1 were assayed by real-time fluorescence quantitative PCR. Results: After continuously cultured for 21days with GSH, Toluidine blue staining and immunofluorescence reaction were all positive in basic induction medium group (group B), basic induction medium+0.5% dimethylsulfoxide (DMSO) group (group BD) and basic induction medium+0.5% DMSO+500μmol/L GSH group (group BDG). Moreover, compared with in group B and group BD, the content of COL2A1 and GAG in group BDG relatively increased and the mRNA expression level of COL2A1 and GAG also comparatively increased(P＜0.05), and both had a significant statistical significance(P＜0.05). Conclusion: GSH might promote the chondrogenic differentiation of hUC-MSCs.
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Received: 19 December 2012
Published: 25 March 2013
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[1] Gupta P K, Das A K, Chullikana A, et al. Mesenchymal stem cells for cartilage repair in osteoarthritis. Stem Cell Res Ther, 2012, 3(4): 25.
[2] Bian L, Zhai D Y, Mauck R L, et al. Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage. Tissue Eng Part A, 2011, 17(7-8): 1137-1145.
[3] Ando W, Fujie H, Moriguchi Y, et al. Detection of abnormalities in the superficial zone of cartilage repaired using a tissue engineered construct derived from synovial stem cells. Eur Cell Mater, 2012, 28(24): 292-307.
[4] Secco M, Zucconi E, Vieira N M. Mesenchymal stem cells from umbilical cord: Do not discard the cord! Neuromuscular Disorders, 2008, 18(1): 17-18.
[5] Wang H S, Hung S C, Peng S T, et al. Mesenchymal stem cells in the Wharton’s Jelly of the human umbilical cord. Stem Cells, 2004, 22(7): 1330-1337.
[6] Qiao C, Xu W, Zhu W, et al. Human mesenchymal stem cells isolated from the umbilical cord. Cell Biol Int, 2008, 32(1): 8-15.
[7] Greish S, Abogresha N, Abdel-Hady Z, et al. Human umbilical cord mesenchymal stem cells as treatment of adjuvant rheumatoid arthritis in a rat model. Stem Cells, 2012, 4(10): 101-109.
[8] Majore I, Moretti P, Stahl F, et al. Growth and differentiation properties of mesenchymal stromal cell populations derived from whole human umbilical cord. Stem Cell Rev, 2011, 7(1): 17-31.
[9] Ruiz-Litago F, Seco J, Echevarría E, et al. Adaptive response in the antioxidant defence system in the course and outcome in first-episode schizophrenia patients: A 12-months follow-up study. Psychiatry Res, 2012, 200(2-3): 218-222.
[10] Carlo M D, Loeser R F. Increased oxidative stress with aging reduces chondrocyte survival: correlation with intracellular glutathione levels. Arthritis Rheum, 2003, 48(12): 3419-3430.
[11] Hatori Y, Clasen S, Hasan N M,et al. Functional partnership of the copper export machinery and glutathione balance in human cells. J Biol Chem, 2012, 287(32): 26678-26687.
[12] Li ZH, Zhao W H, Zhou Q L, et al. Experimental study of velvet antler polypeptides against oxidative damage of osteoarthritis cartilage cells. Zhongguo GU Shang, 2011, 24(3): 245-248.
[13] Takarada-Iemata M, Takarada T, Nakamura Y, et al. Glutamate preferentially suppresses osteoblastogenesis than adipogenesis through the cystine/glutamate antiporter in mesenchymal stem cells. J Cell Physiol, 2011, 226(3): 652-665.
[14] Bjrnsson S. Simultaneous preparation and quantitation of proteoglycans by precipitation with alcian blue. Anal Biochem, 1993, 210(2): 282-291.
[15] 柳菁,宇丽,许超等. 长期培养人脐带间充质干细胞分化能力改变的研究. 中国组织工程杂志,2012, 32(2):8-16. Liu jing, Yu li, Xu chao, et al. Study on differentiation potency changes of long-term cultured human umbilical cord MSCs. China Biotechnology, 2012, 32(2): 8-16.
[16] Tichopad A, Dilger M, Schwarz G, et al. Standardized determination of real-time PCR efficiency from a single reaction set-up. Nucleic Acids Res, 2003, 31(22): 6688.
[17] Cui L, Zhou X, Li J,. et al. Dynamic microRNA Profiles of Hepatic Differentiated Human Umbilical Cord Lining-Derived Mesenchymal Stem Cells. PLoS One, 2012, 7(9): 1-11.
[18] Burra P, Arcidiacono D, Bizzaro D, 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): 88.
[19] Fang T C, Pang C Y, Chiu S C, et al. Renoprotective effect of human umbilical cord-derived mesenchymal stem cells in immunodeficient mice suffering from acute kidney injury. PLoS One, 2012, 7(9): 1-15.
[20] Margossian T, Reppel L, Makdissy N, et al. Mesenchymal stem cells derived from Wharton's jelly: Comparative phenotype analysis between tissue and in vitro expansion. Biomed Mater Eng, 2012, 22(4): 243-254.
[21] Wang L, Detamore M S. Insulin-like growth factor-I improves chondrogenesis of predifferentiated human umbilical cord mesenchymal stromal cells. J Orthop Res, 2009, 27 (8): 1109-1115.
[22] Kuo S M, Chiang M Y, Lan C W, et al. Evaluation of nanoarchitectured collagen type II molecules on cartilage engineering. J Biomed Mater Res A, 2012, 30: 1-10.
[23] Kotwal N, Li J, Sandy J, et al. Initial application of EPIC-μCT to assess mouse articular cartilage morphology and composition: effects of aging and treadmill running. Osteoarthritis Cartilage, 2012, 20(8): 887-895.
[24] Zitka O, Skalickova S, Gumulec J, et al. Redox status expressed as GSH: GSSG ratio as a marker for oxidative stress in paediatric tumour patients. Oncol Lett, 2012, 4(6): 1247-1253.
[25] Liu D, Wang F, Wang Q, et al. Association of glutathione S-transferase M1 polymorphisms and lung cancer risk in a Chinese population. Clin Chim Acta, 2012, 414: 188-190.
[26] Huang H, Yao H, Liu J Y, et al. Development of pyrethroid-like fluorescent substrates for glutathione S-transferase. Anal Biochem, 2012, 431(2): 77-83.
[27] Jocelyne H, Neil W R, Thomas W M. Glutathione, glutathione S-transferase, and glutathione conjugates, complementary markers of oxidative stress in aquatic biota. Environ Sci Pollut Res Int, 2012, 19(6): 2007-2023.
[28] Liu H, Bian W, Liu S, Huang K. Selenium Protects Bone Marrow Stromal Cells Against Hydrogen Peroxide-Induced Inhibition of Osteoblastic Differentiation by Suppressing Oxidative Stress and ERK Signaling Pathway. Biol Trace Elem Res, 2012, 150(1-3): 441-450. |
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