Long-term culture in vitro impairs the immunosuppressive activity of mesenchymal stem cells on T cells

Li,Xue-Yi; Ding,Jin; Zheng,Zhao-Hui; Li,Xiao-Yan; Wu,Zhen-Biao; Zhu,Ping

doi:10.3892/mmr.2012.1039

Long-term culture in vitro impairs the immunosuppressive activity of mesenchymal stem cells on T cells

Authors:
- Xue-Yi Li
- Jin Ding
- Zhao-Hui Zheng
- Xiao-Yan Li
- Zhen-Biao Wu
- Ping Zhu
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Affiliations: Department of Clinical Immunology, State Key Discipline of Cell Biology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Endocrine and Metabolic Diseases, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710068, P.R. China
Published online on: August 17, 2012 https://doi.org/10.3892/mmr.2012.1039
Pages: 1183-1189

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Abstract

Improved knowledge of the immunological properties of mesenchymal stem cells (MSCs) creates a potential cell-mediated immunotherapeutic approach for arthritic diseases. The low frequency of MSCs necessitates their in vitro expansion prior to clinical use. As sequential passage has been used as the most popular strategy for expansion of MSCs, the effect of long-term culture on the immunological properties of MSCs is not clear. In this study, we observed that the morphology of MSCs showed the typical characteristics of the Hayflick model of cellular aging during sequential expansion. The growth kinetics of MSCs decreased while the number of MSCs staining positive for SA β-gal (senescence marker) increased in long-term culture. Although long-term culture exerts less of an effect on the immunophenotype of MSCs, the immunosuppressive effects of MSCs on the allogeneic T-cell proliferation, activation-antigen expression (CD69 and CD25) and cytokine production (IFN-γ, TNF-α, IL-10) were significantly impaired following stimulation with phytohemagglutinin (PHA).

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1	Caplan AI: Mesenchymal stem cells. J Orthop Res. 9:641–650. 1991. View Article : Google Scholar : PubMed/NCBI
2	Deans RJ and Moseley AB: Mesenchymal stem cells: biology and potential clinical uses. Exp Hematol. 28:875–884. 2000. View Article : Google Scholar : PubMed/NCBI
3	Pittenger MF, Mackay AM, Beck SC, et al: Multilineage potential of adult human mesenchymal stem cells. Science. 284:143–147. 1999. View Article : Google Scholar : PubMed/NCBI
4	Ferrari G, Cusella-De Angelis G, Coletta M, et al: Muscle regeneration by bone marrow-derived myogenic progenitors. Science. 279:1528–1530. 1998. View Article : Google Scholar : PubMed/NCBI
5	Liechty KW, MacKenzie TC, Shaaban AF, et al: Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep. Nat Med. 6:1282–1286. 2000. View Article : Google Scholar : PubMed/NCBI
6	Pereira RF, O’Hara MD, Laptev AV, et al: Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta. Proc Natl Acad Sci USA. 95:1142–1147. 1998.PubMed/NCBI
7	Di Nicola M, Carlo-Stella C, Magni M, et al: Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood. 99:3838–3843. 2002.
8	Krampera M, Glennie S, Dyson J, et al: Bone marrow mesenchymal stem cells inhibit the response of naive and memory antigen-specific T cells to their cognate peptide. Blood. 101:3722–3729. 2003. View Article : Google Scholar : PubMed/NCBI
9	Aggarwal S and Pittenger MF: Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood. 105:1815–1822. 2005. View Article : Google Scholar : PubMed/NCBI
10	Bartholomew A, Sturgeon C, Siatskas M, et al: Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. Exp Hematol. 30:42–48. 2002. View Article : Google Scholar : PubMed/NCBI
11	Le Blanc K, Rasmusson I, Sundberg B, et al: Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells. Lancet. 363:1439–1441. 2004.PubMed/NCBI
12	Zappia E, Casazza S, Pedemonte E, et al: Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy. Blood. 106:1755–1761. 2005. View Article : Google Scholar : PubMed/NCBI
13	Zheng ZH, Li XY, Ding J, Jia JF and Zhu P: Allogeneic mesenchymal stem cell and mesenchymal stem cell-differentiated chondrocyte suppress the responses of type II collagen-reactive T cells in rheumatoid arthritis. Rheumatology (Oxford). 47:22–30. 2008. View Article : Google Scholar : PubMed/NCBI
14	Chen FH and Tuan RS: Mesenchymal stem cells in arthritic diseases. Arthritis Res Ther. 10:2232008. View Article : Google Scholar : PubMed/NCBI
15	Baxter MA, Wynn RF, Jowitt SN, Wraith JE, Fairbairn LJ and Bellantuono I: Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion. Stem Cells. 22:675–682. 2004. View Article : Google Scholar : PubMed/NCBI
16	Bonab MM, Alimoghaddam K, Talebian F, Ghaffari SH, Ghavamzadeh A and Nikbin B: Aging of mesenchymal stem cell in vitro. BMC Cell Biol. 71:42006.
17	Weyand CM and Goronzy JJ: Stem cell aging and autoimmunity in rheumatoid arthritis. Trends Mol Med. 10:426–433. 2004. View Article : Google Scholar : PubMed/NCBI
18	Morimoto D, Kuroda S, Kizawa T, et al: Equivalent osteoblastic differentiation function of human mesenchymal stem cells from rheumatoid arthritis in comparison with osteoarthritis. Rheumatology (Oxford). 48:643–649. 2009. View Article : Google Scholar
19	Kassem M, Ankersen L, Eriksen EF, Clark BF and Rattan SI: Demonstration of cellular aging and senescence in serially passaged long-term cultures of human trabecular osteoblasts. Osteoporos Int. 7:514–524. 1997. View Article : Google Scholar : PubMed/NCBI
20	Dimri GP, Lee X, Basile G, et al: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA. 92:9363–9367. 1995. View Article : Google Scholar : PubMed/NCBI
21	Groh ME, Maitra B, Szekely E and Koc ON: Human mesenchymal stem cells require monocyte-mediated activation to suppress alloreactive T cells. Exp Hematol. 33:928–934. 2005. View Article : Google Scholar : PubMed/NCBI
22	Le Blanc K, Rasmusson I, Gotherstrom C, et al: Mesenchymal stem cells inhibit the expression of CD25 (interleukin-2 receptor) and CD38 on phytohaemagglutinin-activated lymphocytes. Scand J Immunol. 60:307–315. 2004.PubMed/NCBI
23	Zhu P, Li XY, Wang HK, et al: Oral administration of type-II collagen peptide 250–270 suppresses specific cellular and humoral immune response in collagen-induced arthritis. Clin Immunol. 122:75–84. 2007.
24	Fridenshtein A, Petrakova KV, Kuralesova AI and Frolova GI: Precursor cells for osteogenic and hemopoietic tissues. Analysis of heterotopic transplants of bone marrow. Tsitologiia. 10:557–567. 1968.PubMed/NCBI
25	Lu L, Zhao C, Liu Y, et al: Therapeutic benefit of TH-engineered mesenchymal stem cells for Parkinson’s disease. Brain Res. 15:46–51. 2005.PubMed/NCBI
26	Studeny M, Marini FC, Dembinski JL, et al: Mesenchymal stem cells: potential precursors for tumor stroma and targeted-delivery vehicles for anticancer agents. J Natl Cancer Inst. 96:1593–1603. 2004. View Article : Google Scholar : PubMed/NCBI
27	Friedenstein AJ, Deriglasova UF, Kulagina NN, et al: Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method. Exp Hematol. 2:83–92. 1974.PubMed/NCBI
28	Dressler MR, Butler DL and Boivin GP: Effects of age on the repair ability of mesenchymal stem cells in rabbit tendon. J Orthop Res. 23:287–293. 2005. View Article : Google Scholar : PubMed/NCBI
29	Liu L, DiGirolamo CM, Navarro PA, Blasco MA and Keefe DL: Telomerase deficiency impairs differentiation of mesenchymal stem cells. Exp Cell Res. 294:1–8. 2004. View Article : Google Scholar : PubMed/NCBI
30	Stolzing A and Scutt A: Age-related impairment of mesenchymal progenitor cell function. Aging Cell. 5:213–224. 2006. View Article : Google Scholar : PubMed/NCBI
31	Zheng H, Martin JA, Duwayri Y, Falcon G and Buckwalter JA: Impact of aging on rat bone marrow-derived stem cell chondrogenesis. J Gerontol A Biol Sci Med Sci. 62:136–148. 2007. View Article : Google Scholar : PubMed/NCBI
32	Carrancio S, Lopez-Holgado N, Sanchez-Guijo FM, et al: Optimization of mesenchymal stem cell expansion procedures by cell separation and culture conditions modification. Exp Hematol. 36:1014–1021. 2008. View Article : Google Scholar : PubMed/NCBI
33	Zohar R, Sodek J and McCulloch CA: Characterization of stromal progenitor cells enriched by flow cytometry. Blood. 90:3471–3481. 1997.PubMed/NCBI
34	Matsubara T, Tsutsumi S, Pan H, et al: A new technique to expand human mesenchymal stem cells using basement membrane extracellular matrix. Biochem Biophys Res Commun. 313:503–508. 2004. View Article : Google Scholar : PubMed/NCBI
35	Schallmoser K, Rohde E, Reinisch A, et al: Rapid large-scale expansion of functional mesenchymal stem cells from unmanipulated bone marrow without animal serum. Tissue engineering. 14:185–196. 2008. View Article : Google Scholar : PubMed/NCBI
36	Hung SC, Yang DM, Chang CF, et al: Immortalization without neoplastic transformation of human mesenchymal stem cells by transduction with HPV16 E6/E7 genes. Int J Cancer. 110:313–319. 2004. View Article : Google Scholar : PubMed/NCBI
37	Shi S, Gronthos S, Chen S, et al: Bone formation by human postnatal bone marrow stromal stem cells is enhanced by telomerase expression. Nat Biotechnol. 20:587–591. 2002. View Article : Google Scholar : PubMed/NCBI