Abstract
Cellular therapies are becoming a major focus for the treatment of demyelinating diseases such as multiple sclerosis (MS), therefore it is important to identify the most effective cell types that promote myelin repair. Several components contribute to the relative benefits of specific cell types including the overall efficacy of the cell therapy, the reproducibility of treatment, the mechanisms of action of distinct cell types and the ease of isolation and generation of therapeutic populations. A range of distinct cell populations promote functional recovery in animal models of MS including neural stem cells and mesenchymal stem cells derived from different tissues. Each of these cell populations has advantages and disadvantages and likely works through distinct mechanisms. The relevance of such mechanisms to myelin repair in the adult central nervous system is unclear since the therapeutic cells are generally derived from developing animals. Here we describe the isolation and characterization of a population of neural cells from the adult spinal cord that are characterized by the expression of the cell surface glycoprotein NG2. In functional studies, injection of adult NG2+ cells into mice with ongoing MOG35–55-induced experimental autoimmune encephalomyelitis (EAE) enhanced remyelination in the CNS while the number of CD3+ T cells in areas of spinal cord demyelination was reduced approximately three-fold. In vivo studies indicated that in EAE, NG2+ cells stimulated endogenous repair while in vitro they responded to signals in areas of induced inflammation by differentiating into oligodendrocytes. These results suggested that adult NG2+ cells represent a useful cell population for promoting neural repair in a variety of different conditions including demyelinating diseases such as MS.
Keywords: NG2 glycoprotein, myelin, oligodendrocytes, experimental autoimmune encephalomyelitis, multiple sclerosis, remyelination
Contributor Information
Lianhua Bai, Email: xiaohua42@hotmail.com.
Robert H. Miller, Email: robert.miller@case.edu
References
- [1].Miller RH. The promise of stem cells for neural repair. Brain Res. 2006;1091:258–264. doi: 10.1016/j.brainres.2006.01.073. [DOI] [PubMed] [Google Scholar]
- [2].Miller RH, Bai L, Lennon DP, Caplan AI. The potential of mesenchymal stem cells for neural repair. Discov Med. 2010;9:236–242. [PubMed] [Google Scholar]
- [3].Bai L, Lennon DP, Eaton V, Maier K, Caplan AI, Miller SD, et al. Human bone marrow-derived mesenchymal stem cells induce Th2-polarized immune response and promote endogenous repair in animal models of multiple sclerosis. Glia. 2009;57:1192–1203. doi: 10.1002/glia.20841. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [4].Sharp J, Keirstead HS. Stem cell-based cell replacement strategies for the central nervous system. Neurosci Lett. 2009;456:107–111. doi: 10.1016/j.neulet.2008.04.106. [DOI] [PubMed] [Google Scholar]
- [5].Auletta JJ, Bartholomew AM, Maziarz RT, Deans RJ, Miller RH, Lazarus HM, et al. The potential of mesenchymal stromal cells as a novel cellular therapy for multiple sclerosis. Immunotherapy. 2012;4:529–547. doi: 10.2217/imt.12.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [6].Bai L, Lennon DP, Caplan AI, DeChant A, Hecker J, Kranso J, et al. Hepatocyte growth factor mediates mesenchymal stem cell-induced recovery in multiple sclerosis models. Nat Neurosci. 2012;15:862–870. doi: 10.1038/nn.3109. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [7].Ben-Hur T. Cell therapy for multiple sclerosis. Neurotherapeutics. 2011;8:625–642. doi: 10.1007/s13311-011-0073-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [8].Pluchino S, Quattrini A, Brambilla E, Gritti A, Salani G, Dina G, et al. Injection of adult neurospheres induces recovery in a chronic model of multiple sclerosis. Nature. 2003;422:688–694. doi: 10.1038/nature01552. [DOI] [PubMed] [Google Scholar]
- [9].Bassi G, Pacelli L, Carusone R, Zanoncello J, Krampera M. Adipose-derived stromal cells (ASCs) Transfus Apher Sci. 2012;47:193–198. doi: 10.1016/j.transci.2012.06.004. [DOI] [PubMed] [Google Scholar]
- [10].Fouraschen SM, Pan Q, de Ruiter PE, Farid WR, Kazemier G, Kwekkeboom J, et al. Secreted factors of human liver-derived mesenchymal stem cells promote liver regeneration early after partial hepatectomy. Stem Cells Dev. 2012;21:2410–2419. doi: 10.1089/scd.2011.0560. [DOI] [PubMed] [Google Scholar]
- [11].Hoffman AM, Paxson JA, Mazan MR, Davis AM, Tyagi S, Murthy S, et al. Lung-derived mesenchymal stromal cell post-transplantation survival, persistence, paracrine expression, and repair of elastase-injured lung. Stem Cells Dev. 2011;20:1779–1792. doi: 10.1089/scd.2011.0105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [12].Huang Y, Chen P, Zhang CB, Ko GJ, Ruiz M, Fiorina P, et al. Kidney-derived mesenchymal stromal cells modulate dendritic cell function to suppress alloimmune responses and delay allograft rejection. Transplantation. 2010;90:1307–1311. doi: 10.1097/TP.0b013e3181fdd9eb. [DOI] [PubMed] [Google Scholar]
- [13].Caplan AI. All MSCs are pericytes? Cell Stem Cell. 2008;3:229–230. doi: 10.1016/j.stem.2008.08.008. [DOI] [PubMed] [Google Scholar]
- [14].Vaculik C, Schuster C, Bauer W, Iram N, Pfisterer K, Kramer G, et al. Human dermis harbors distinct mesenchymal stromal cell subsets. J Invest Dermatol. 2012;132:563–574. doi: 10.1038/jid.2011.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [15].Caplan AI. New era of cell-based orthopedic therapies. Tissue Eng Part B Rev. 2009;15:195–200. doi: 10.1089/ten.teb.2008.0515. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [16].Caplan AI, Correa D. PDGF in bone formation and regeneration: new insights into a novel mechanism involving MSCs. J Orthop Res. 2011;29:1795–1803. doi: 10.1002/jor.21462. [DOI] [PubMed] [Google Scholar]
- [17].Dore-Duffy P, Mehedi A, Wang X, Bradley M, Trotter R, Gow A. Immortalized CNS pericytes are quiescent smooth muscle actin-negative and pluripotent. Microvasc Res. 2011;82:18–27. doi: 10.1016/j.mvr.2011.04.003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [18].Canfield AE, Sutton AB, Hoyland JA, Schor AM. Association of thrombospondin-1 with osteogenic differentiation of retinal pericytes in vitro. J Cell Sci. 1996;109(Pt2):343–353. doi: 10.1242/jcs.109.2.343. [DOI] [PubMed] [Google Scholar]
- [19].Doherty MJ, Ashton BA, Walsh S, Beresford JN, Grant ME, Canfield AE. Vascular pericytes express osteogenic potential in vitro and in vivo. J Bone Miner Res. 1998;13:828–838. doi: 10.1359/jbmr.1998.13.5.828. [DOI] [PubMed] [Google Scholar]
- [20].Dore-Duffy P. Pericytes: pluripotent cells of the blood brain barrier. Curr Pharm Des. 2008;14:1581–1593. doi: 10.2174/138161208784705469. [DOI] [PubMed] [Google Scholar]
- [21].Hirschi KK, D’Amore PA. Pericytes in the microvasculature. Cardiovasc Res. 1996;32:687–698. [PubMed] [Google Scholar]
- [22].Lundberg C, Martinez-Serrano A, Cattaneo E, McKay RD, Bjorklund A. Survival, integration, and differentiation of neural stem cell lines after transplantation to the adult rat striatum. Exp Neurol. 1997;145:342–360. doi: 10.1006/exnr.1997.6503. [DOI] [PubMed] [Google Scholar]
- [23].Ozerdem U, Stallcup WB. Pathological angiogenesis is reduced by targeting pericytes via the NG2 proteoglycan. Angiogenesis. 2004;7:269–276. doi: 10.1007/s10456-004-4182-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [24].Stallcup WB. The NG2 proteoglycan: past insights and future prospects. J Neurocytol. 2002;31:423–435. doi: 10.1023/A:1025731428581. [DOI] [PubMed] [Google Scholar]
- [25].Nishiyama A. Polydendrocytes: NG2 cells with many roles in development and repair of the CNS. Neuroscientist. 2007;13:62–76. doi: 10.1177/1073858406295586. [DOI] [PubMed] [Google Scholar]
- [26].Stallcup WB. The NG2 antigen, a putative lineage marker: immunofluorescent localization in primary cultures of rat brain. Dev Biol. 1981;83:154–165. doi: 10.1016/S0012-1606(81)80018-8. [DOI] [PubMed] [Google Scholar]
- [27].Zhu X, Hill RA, Nishiyama A. NG2 cells generate oligodendrocytes and gray matter astrocytes in the spinal cord. Neuron Glia Biol. 2008;4:19–26. doi: 10.1017/S1740925X09000015. [DOI] [PubMed] [Google Scholar]
- [28].Nishiyama A, Komitova M, Suzuki R, Zhu X. Polydendrocytes (NG2 cells): multifunctional cells with lineage plasticity. Nat Rev Neurosci. 2009;10:9–22. doi: 10.1038/nrn2495. [DOI] [PubMed] [Google Scholar]
- [29].Karadottir R, Hamilton NB, Bakiri Y, Attwell D. Spiking and nonspiking classes of oligodendrocyte precursor glia in CNS white matter. Nat Neurosci. 2008;11:450–456. doi: 10.1038/nn2060. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [30].MacFadyen J, Savage K, Wienke D, Isacke CM. Endosialin is expressed on stromal fibroblasts and CNS pericytes in mouse embryos and is downregulated during development. Gene Expr Patterns. 2007;7:363–369. doi: 10.1016/j.modgep.2006.07.006. [DOI] [PubMed] [Google Scholar]
- [31].Shepro D, Morel NM. Pericyte physiology. FASEB J. 1993;7:1031–1038. doi: 10.1096/fasebj.7.11.8370472. [DOI] [PubMed] [Google Scholar]
- [32].Edelman DA, Jiang Y, Tyburski J, Wilson RF, Steffes C. Pericytes and their role in microvasculature homeostasis. J Surg Res. 2006;135:305–311. doi: 10.1016/j.jss.2006.06.010. [DOI] [PubMed] [Google Scholar]
- [33].von Tell D, Armulik A, Betsholtz C. Pericytes and vascular stability. Exp Cell Res. 2006;312:623–629. doi: 10.1016/j.yexcr.2005.10.019. [DOI] [PubMed] [Google Scholar]
- [34].Balabanov R, Dore-Duffy P. Role of the CNS microvascular pericyte in the blood-brain barrier. J Neurosci Res. 1998;53:637–644. doi: 10.1002/(SICI)1097-4547(19980915)53:6<637::AID-JNR1>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
- [35].Ballabh P, Braun A, Nedergaard M. The blood-brain barrier: an overview: structure, regulation, and clinical implications. Neurobiol Dis. 2004;16:1–13. doi: 10.1016/j.nbd.2003.12.016. [DOI] [PubMed] [Google Scholar]
- [36].Dore-Duffy P, Katychev A, Wang X, Van Buren E. CNS microvascular pericytes exhibit multipotential stem cell activity. J Cereb Blood Flow Metab. 2006;26:613–624. doi: 10.1038/sj.jcbfm.9600272. [DOI] [PubMed] [Google Scholar]
- [37].Wight PA, Duchala CS, Shick HE, Gudz TI, Macklin WB. Expression of a myelin proteolipid protein (Plp)-lacZ transgene is reduced in both the CNS and PNS of Plp(jp) mice. Neurochem Res. 2007;32:343–351. doi: 10.1007/s11064-006-9202-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [38].Garbelli R, Zucca I, Milesi G, Mastropietro A, D’Incerti L, Tassi L, et al. Combined 7-T MRI and histopathologic study of normal and dysplastic samples from patients with TLE. Neurology. 2011;76:1177–1185. doi: 10.1212/WNL.0b013e318212aae1. [DOI] [PubMed] [Google Scholar]
- [39].Sun HS, French RJ, Feng ZP. A method for identifying viable and damaged neurons in adult mouse brain slices. Acta Histochem. 2009;111:531–537. doi: 10.1016/j.acthis.2008.06.005. [DOI] [PubMed] [Google Scholar]
- [40].Jariyapongskul A, Nakano A, Yamaguchi S, Nageswari K, Niimi H. Maturity of pericytes in cerebral neocapillaries induced by growth factors: fluorescence immuno-histochemical analysis using confocal laser microscopy. Clin Hemorheol Microcirc. 2003;29:417–421. [PubMed] [Google Scholar]
- [41].Sims DE. The pericyte—a review. Tissue Cell. 1986;18:153–174. doi: 10.1016/0040-8166(86)90026-1. [DOI] [PubMed] [Google Scholar]
- [42].Orlidge A, D’Amore PA. Cell specific effects of glycosaminoglycans on the attachment and proliferation of vascular wall components. Microvasc Res. 1986;31:41–53. doi: 10.1016/0026-2862(86)90005-1. [DOI] [PubMed] [Google Scholar]
- [43].Yang Z, Watanabe M, Nishiyama A. Optimization of oligodendrocyte progenitor cell culture method for enhanced survival. J Neurosci Methods. 2005;149:50–56. doi: 10.1016/j.jneumeth.2005.05.003. [DOI] [PubMed] [Google Scholar]
- [44].Kucharova K, Chang Y, Boor A, Yong VW, Stallcup WB. Reduced inflammation accompanies diminished myelin damage and repair in the NG2 null mouse spinal cord. J Neuroinflammation. 2011;8:158. doi: 10.1186/1742-2094-8-158. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [45].Tripathi RB, Rivers LE, Young KM, Jamen F, Richardson WD. NG2 glia generate new oligodendrocytes but few astrocytes in a murine experimental autoimmune encephalomyelitis model of demyelinating disease. J Neurosci. 2010;30:16383–16390. doi: 10.1523/JNEUROSCI.3411-10.2010. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [46].Pringle N, Collarini EJ, Mosley MJ, Heldin CH, Westermark B, Richardson WD. PDGF A chain homodimers drive proliferation of bipotential (O-2A) glial progenitor cells in the developing rat optic nerve. EMBO J. 1989;8:1049–1056. doi: 10.1002/j.1460-2075.1989.tb03472.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [47].Tsai HH, Macklin WB, Miller RH. Distinct modes of migration position oligodendrocyte precursors for localized cell division in the developing spinal cord. J Neurosci Res. 2009;87:3320–3330. doi: 10.1002/jnr.22058. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [48].Kaplan JM, Youd ME, Lodie TA. Immunomodulatory activity of mesenchymal stem cells. Curr Stem Cell Res Ther. 2011;6:297–316. doi: 10.2174/157488811797904353. [DOI] [PubMed] [Google Scholar]
- [49].Payne NL, Sun G, Herszfeld D, Tat-Goh PA, Verma PJ, Parkington HC, et al. Comparative study on the therapeutic potential of neurally differentiated stem cells in a mouse model of multiple sclerosis. PLoS One. 2012;7:e35093. doi: 10.1371/journal.pone.0035093. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [50].Miller RH, Bai L. Translating stem cell therapies to the clinic. Neurosci Lett. 2012;519:87–92. doi: 10.1016/j.neulet.2012.01.043. [DOI] [PubMed] [Google Scholar]
- [51].Busch SA, Horn KP, Cuascut FX, Hawthorne AL, Bai L, Miller RH, et al. Adult NG2+ cells are permissive to neurite outgrowth and stabilize sensory axons during macrophageinduced axonal dieback after spinal cord injury. J Neurosci. 2010;30:255–265. doi: 10.1523/JNEUROSCI.3705-09.2010. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [52].Whitman LM, Blanc CA, Schaumburg CS, Rowitch DH, Lane TE. Olig1 function is required for remyelination potential of transplanted neural progenitor cells in a model of viral-induced demyelination. Exp Neurol. 2012;235:380–387. doi: 10.1016/j.expneurol.2012.03.003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [53].Kondo T, Raff M. Oligodendrocyte precursor cells reprogrammed to become multipotential CNS stem cells. Science. 2000;289:1754–1757. doi: 10.1126/science.289.5485.1754. [DOI] [PubMed] [Google Scholar]
- [54].Nishiyama A, Yang Z, Butt A. Astrocytes and NG2-glia: what’s in a name? J Anat. 2005;207:687–693. doi: 10.1111/j.1469-7580.2005.00489.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [55].Dincman TA, Beare JE, Ohri SS, Whittemore SR. Isolation of cortical mouse oligodendrocyte precursor cells. J Neurosci Methods. 2012;209:219–226. doi: 10.1016/j.jneumeth.2012.06.017. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [56].Staugaitis SM, Trapp BD. NG2-positive glia in the human central nervous system. Neuron Glia Biol. 2009;5:35–44. doi: 10.1017/S1740925X09990342. [DOI] [PubMed] [Google Scholar]
- [57].Carlesi C, Pasquali L, Piazza S, Lo Gerfo A, Caldarazzo Ienco E, Alessi R, et al. Strategies for clinical approach to neurodegeneration in Amyotrophic lateral sclerosis. Arch Ital Biol. 2011;149:151–167. doi: 10.4449/aib.v149i1.1267. [DOI] [PubMed] [Google Scholar]
- [58].Naganska E, Matyja E. Amyotrophic lateral sclerosis — looking for pathogenesis and effective therapy. Folia Neuropathol. 2011;49:1–13. [PubMed] [Google Scholar]
- [59].Lepore AC, Dejea C, Carmen J, Rauck B, Kerr DA, Sofroniew MV, et al. Selective ablation of proliferating astrocytes does not affect disease outcome in either acute or chronic models of motor neuron degeneration. Exp Neurol. 2008;211:423–432. doi: 10.1016/j.expneurol.2008.02.020. [DOI] [PMC free article] [PubMed] [Google Scholar]
- [60].Kang SH, Fukaya M, Yang JK, Rothstein JD, Bergles DE. NG2+ CNS glial progenitors remain committed to the oligodendrocyte lineage in postnatal life and following neurodegeneration. Neuron. 2010;68:668–681. doi: 10.1016/j.neuron.2010.09.009. [DOI] [PMC free article] [PubMed] [Google Scholar]