The effect of reduced glutathione on the chondrogenesis of human umbilical cord mesenchymal stem cells


It has been discussed whether reduced glutathione (GSH) could promote the chondrogenic differentiation ability of human umbilical cord mesenchymal stem cells (hUC-MSCs). hUC-MSCs were isolated from human umbilical cord and their specificity was identified, then induced into cartilage-like cells in chondrogenic induction medium with transforming growth factor beta 1 (TGF-β1), especially with GSH. The morphological change before and after induction was observed through inverted phase contrast microscope, Type II collagen (COL2-A1) and glycosaminoglycan (GAG) were analyzed qualitatively by Toluidine blue and immunofluorescence technique, respectively, the contents of COL2-A1 and GAG were estimated from the determination of hydroxyproline content and Alcian Blue method separately. The mRNA expressions of GAG and COL2-A1 were assayed by real-time fluorescence quantitative PCR. After continuously cultured for 21 days 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 +500 μM GSH group (group BDG). Moreover, compared with group B and group BD, the contents of COL2-A1 and GAG in group BDG relatively increased and the mRNA expression level of COL2-A1 and GAG also comparatively increased (P < 0.05) and both had a significant statistical significance (P < 0.05). So GSH might promote the induction of hUC-MSCs to differentiate into cartilage-like cells.

Share and Cite:

Luo, E. , Zhang, J. , Liu, J. , Yu, L. and Tang, M. (2013) The effect of reduced glutathione on the chondrogenesis of human umbilical cord mesenchymal stem cells. Journal of Biomedical Science and Engineering, 6, 775-781. doi: 10.4236/jbise.2013.68094.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] [1] Keeney, M., Lai, J.H. and Yang, F. (2011) Recent progress in cartilage tissue engineering. Current Opinion in Biotechnology, 22, 734-740. doi:10.1016/j.copbio.2011.04.003
[2] Gupta, P.K., Chullikana, A., et al. (2012) Mesenchymal stem cells for cartilage repair in osteoarthritis. Stem Cell Research & Therapy, 3, 25.
[3] Bian, L., Mauck, R.L., et al. (2011) Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage. Tissue Engineering, Parts A, 17, 1137-1145. doi:10.1089/ten.tea.2010.0531
[4] Ando, W., Moriguchi, Y., et al. (2012) Detection of abnormalities in the superficial zone of cartilage repaired using a tissue engineered construct derived from synovial stem cells. European Cells and Materials, 28, 292-307.
[5] Meirelles, L.D.S. and Nardi, N.B. (2009) Methodology, biology and clinical applications of mesenchymal stem cells. Frontiers in Bioscience, 14, 4281-4298. doi:10.2741/3528
[6] Csaki, C., Schneider, P.R.A. and Shakibaei, M. (2008) Mesenchymal stem cells as a potential pool for cartilage tissue engineering. Annals of Anatomy-Anatomischer Anzeiger, 190, 395-412. doi:10.1016/j.aanat.2008.07.007
[7] Chang, S.C.N., et al. (2008) Comparisons between sources of mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord and bone marrow. Tissue Engineering Part A, 14, 829-829.
[8] Meyer, T., Pfeifroth, A. and Hoecht, B. (2008) Isolation and characterisation of mesenchymal stem cells in Wharton’s jelly of the human umbilical cord: Potent cells for cell-based therapies in paediatric surgery? European Surgery-Acta Chirurgica Austriaca, 40, 239-244. doi:10.1007/s10353-008-0417-x
[9] Wang, H.S., et al. (2004) Mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord. Stem Cells, 22, 1330-1337. doi:10.1634/stemcells.2004-0013
[10] Cui, L., et al. (2012) Dynamic microRNA profiles of hepatic differentiated human umbilical cord lining-derived mesenchymal stem cells. PLoS One, 7. doi:10.1371/journal.pone.0044737
[11] Fang, T.-C., et al. (2012) Renoprotective effect of human umbilical cord-derived mesenchymal stem cells in immunodeficient mice suffering from acute kidney injury. PLoS One, 7. doi:10.1371/journal.pone.0046504
[12] Margossian, T., et al. (2012) Mesenchymal stem cells derived from Wharton’s jelly: Comparative phenotype analysis between tissue and in vitro expansion. Bio-Medical Materials and Engineering, 22, 243-254.
[13] Ruiz-Litago, F., et al. (2012) Adaptive response in the antioxidant defence system in the course and outcome in first-episode schizophrenia patients: A 12-months followup study. Psychiatry Research, 200, 218-222. doi:10.1016/j.psychres.2012.07.024
[14] Zitka, O., et al. (2012) Redox status expressed as GSH: GSSG ratio as a marker for oxidative stress in paediatric tumour patients. Oncology Letters, 4, 1247-1253.
[15] Hatori, Y., et al. (2012) Functional partnership of the copper export machinery and glutathione balance in human cells. Journal of Biological Chemistry, 287, 26678-26687. doi:10.1074/jbc.M112.381178
[16] Liu, H., et al. (2012) Selenium protects bone marrow stromal cells against hydrogen peroxide-induced inhibition of osteoblastic differentiation by suppressing oxidative stress and erk signaling pathway. Biological Trace Element Research, 150, 441-450. doi:10.1007/s12011-012-9488-4
[17] Del Carlo, M. and Loeser, R.F. (2003) Increased oxidative stress with aging reduces chondrocyte survival correlation with intracellular glutathione levels. Arthritis and Rheumatism, 48, 3419-3430. doi:10.1002/art.11338
[18] Li, Z.-H., Zhao, W.-H. and Zhou, Q.-L. (2011) Experimental study of velvet antler polypeptides against oxidative damage of osteoarthritis cartilage cells. China Journal of Orthopaedics and Traumatology, 24, 245-248.
[19] Takarada-Iemata, M., et al. (2011) Glutamate preferentially suppresses osteoblastogenesis than adipogenesis through the cystine/glutamate antiporter in mesenchymal stem cells. Journal of Cellular Physiology, 226, 652-665. doi:10.1002/jcp.22390
[20] Lo, W.-C., et al. (2013) Preferential therapy for osteoarthritis by cord blood MSCs through regulation of chondrogenic cytokines. Biomaterials, 34, 4739-4748. doi:10.1016/j.biomaterials.2013.03.016
[21] Bjornsson, S. (1993) Simultaneous preparation and quantitation of proteoglycans by precipitation with alcian blue. Analytical Biochemistry, 210, 282-291. doi:10.1006/abio.1993.1197
[22] Tichopad, A., et al. (2003) Standardized determination of real-time PCR efficiency from a single reaction set-up. Nucleic Acids Research, 31. doi:10.1093/nar/gng122
[23] Kotwal, N., Sandy, J., et al. (2012) Initial application of EPIC-μCT to assess mouse articular cartilage morphology and composition: Effects of aging and treadmill running. Osteoarthritis Cartilage, 20, 887-895. doi:10.1016/j.joca.2012.04.012
[24] Kuo, S.M., et al. (2013) Evaluation of nanoarchitectured collagen type II molecules on cartilage engineering. Journal of Biomedical Materials Research Part A, 101, 368-377.
[25] Gupta, P.K., et al. (2012) Mesenchymal stem cells for cartilage repair in osteoarthritis. Stem Cell Research & Therapy, 3. doi:10.1002/jbm.a.34335

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.