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NT-3 gene modified Schwann cells promote TrkC gene modified mesenchymal stem cells to differentiate into neuron-like cells in vitro

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Abstract

Reports of neuronal differentiation of bone marrow derived mesenchymal stem cells (MSCs) suggested the possibility that these cells could serve as a source of treatment for spinal cord injury. However, the percentages of neuron-like cells differentiated from the MSCs were relatively low both in vitro and in vivo. Here, we investigated whether co-culture of human neurotrophin-3 (NT-3) gene modified Schwann cells (SCs) and human NT-3 receptor tyrosine protein kinase C (TrkC) gene modified MSCs could increase differentiation of neuron-like cells from MSCs. It was shown that MSCs were significantly promoted to differentiate into neuron-like cells, as evidenced immunocytochemically by the expression of neuronal markers, including nestin, β-III-tubulin, MAP2 and PSD95, 7 days after co-culture. However, the expression of glial fibrillary acidic protein (GFAP)—an astrocyte marker in these cells—was not so obvious. These results demonstrate that the binding of overexpressed NT-3 in SCs and its receptor TrkC in MSCs can be considered to stimulate the increased rate of neuronal differentiation.

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References

  • Abouelfetouh A, Kondoh T, Ehara K, Kohmura E (2004) Morphological differentiation of bone marrow stromal cells into neuron-like cells after co-culture with hippocampal slice. Brain Res 1029:114–119

    Article  CAS  PubMed  Google Scholar 

  • Axanchi R, Bernal G, Gupta R, Keirstead HS (2004) Combined demyelination plus Schwann cell transplantation therapy increases spread of cells and axonal regeneration following contusion injury. J Neurotrauma 21:775–788

    Article  Google Scholar 

  • Blesch A, Lu P, Tuszynski MH (2002) Neurotrophic factors, gene therapy, and neural stem cells for spinal cord repair. Brain Res Bull 57:833–838

    Article  CAS  PubMed  Google Scholar 

  • El-Husseini AE, Schnell E, Chetkorich DM, Nicoll RA, Bredt DS (2000) PSD-95 involvement in maturation of excitatory synapses. Science 290:1304–1368

    Article  Google Scholar 

  • Hofstetter CP, Schwarz EJ, Hess D et al (2002) Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc Natl Acad Sci USA 99:2199–2204

    Article  CAS  PubMed  Google Scholar 

  • Jin K, Mao XO, Batteur S, Sun Y, Greenberg DA (2003) Induction of neuronal markers in bone marrow cells: differential effects of growth factors and patterns of intracellular expression. Exp Neurol 184:78–89

    Article  CAS  PubMed  Google Scholar 

  • Jori FP, Napolitano MA, Melone MA et al (2005) Molecular pathways involved in neural in vitro differentiation of marrow stromal stem cells. J Cell Biochem 94:645–655

    Article  CAS  PubMed  Google Scholar 

  • Klopotowska D, Strzadala L (2005) The role of TrkC receptor and neurotrophin-3 in the development and function of neural cells. Postepy Hig Med Dosw 59:517–522

    Google Scholar 

  • Kopen GC, Prckop DJ, Phinney DG (1999) Marrow stromal cells migrate throughout forebrain and cerebellum and diffentiate into astrocytes after injection into neonatal mouse brain. Proc Natl Acad Sci USA 96:10711–10716

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Chen J, Chen XG et al (2002) Human marrow stromal cell therapy for stroke in rat: neurotrophins and functional recovery. Neurology 59:514–523

    CAS  PubMed  Google Scholar 

  • Qian L, Saltzman WM (2004) Improving the expansion and neuronal differentiation of mesenchymal stem cells through culture surface modification. Biomaterials 25:1331–1337

    Article  CAS  PubMed  Google Scholar 

  • Sanchez-Ramos J, Song S, Cardozo-Pelaez F (2000) Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol 164:247–256

    Article  CAS  PubMed  Google Scholar 

  • Shetty AK, Turner DA (1998) In vitro survival and differentiation of neurons derived from epidermal growth factor-responsive postnatal hippocampal stem cells: inducing effects of brain-derived neurotrophic factor. J Neurobiol 35:395–425

    Article  CAS  PubMed  Google Scholar 

  • Ullian EM, Harris BT, Wu A, Chan JR, Barres BA (2004) Schwann cells and astrocytes induce synapse formation by spinal motor neurons in culture. Mol Cell Neurosci 25:241–251

    Article  CAS  PubMed  Google Scholar 

  • Wang JM, Zeng YS, Liu RY, Wu LZ, Xue G, Huang WL, Xiong Y, Wang YH (2005) Construction of a recombinant adenovirus vector expression human neurotrophin-3 with biological activity (in Chinese). Acta Anat Sin 36:501–506

    Google Scholar 

  • Wang JM, Zeng YS, Liu RY, Huang WL, Xiong Y, Wang YH, Chen SJ, Teng YD (2007) Recombinant adenovirus vector-mediated functional expression of neurotropin-3 receptor (TrkC) in neural stem cells. Exp Neurol 203:123–127

    Article  CAS  PubMed  Google Scholar 

  • Wang TT, Tio M, Lee W, Beerheide W, Udolph G (2007) Neural differentiation of mesenchymal-like stem cells from cord blood is mediated by PKA. Biochem Biophys Res Commun 357:1021–1027

    Article  CAS  PubMed  Google Scholar 

  • Woodbury D, Schwarz EJ, Prockop DJ, Black IB (2000) Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 61:364–370

    Article  CAS  PubMed  Google Scholar 

  • Woodbury D, Reynolds K, Black IB (2002) Adult bone marrow stromal stem cells express germline, ectodermal, endodermal, and mesodermal genes prior to neurogenesis. J Neurosci Res 69:908–917

    Article  CAS  PubMed  Google Scholar 

  • Wu J, Sun TS, Ren JX, Wang XZ (2008) Ex vivo non-viral vector-mediated neurotrophin-3 gene transfer to olfactory ensheathing glia: effects on axonal regeneration and functional recovery after implantation in rats with spinal cord injury. Neurosci Bull 24:57–65

    Article  PubMed  Google Scholar 

  • Xu XM, Chen A, Guénard V, Kleitman N, Bunge MB (1997) Bridging Schwann cell transplants promote axonal regeneration from both the rostral and caudal stumps of transected adult rat spinal cord. J Neurocytol 26:1–16

    Article  CAS  PubMed  Google Scholar 

  • Yano S, Kuroda S, Lee JB et al (2005) In vivo fluorescence tracking of bone marrow stromal cells transplanted into a pneumatic injury model of rat spinal cord. J Neurotrauma 22:907–918

    Article  PubMed  Google Scholar 

  • Zhang W, Zeng YS, Zhang XB, Wang JM, Zhang W, Chen SJ (2006) Combination of adenoviral vector-mediated neurotrophin-3 gene transfer and retinoic acid promotes adult bone marrow cells to differentiate into neuronal phenotypes. Neurosci Lett 408:98–103

    Article  CAS  PubMed  Google Scholar 

  • Zurita M, Vaquero J (2006) Bone marrow stromal cells can achieve cure of chronic paraplegic rats: functional and morphological outcome one year after transplantation. Neurosci Lett 402:51–56

    Article  CAS  PubMed  Google Scholar 

  • Zurita M, Vaquero J, Oya S, Miguel M (2005) Schwann cells induce neuronal differentiation of bone marrow stromal cells. Neuroreport 16:505–508

    Article  PubMed  Google Scholar 

  • Zurita M, Vaquero J, Oya S, Bonilla C, Aguayo C (2007) Neurotrophic Schwann-cell factors induce neural differentiation of bone marrow stromal cells. Neuroreport 18:1713–1717

    Article  PubMed  Google Scholar 

  • Zurita M, Bonilla C, Otero L, Aguayo C, Vaquero J (2008) Neural transdifferentiation of bone marrow stromal cells obtained by chemical agents is a short-time reversible phenomenon. Neurosci Res 60:275–280

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This research was co-supported by the Doctor Subject Points Foundation of Chinese Education Ministry (DSPFCEM 2004558068) and the Chinese National Natural Science Foundation (CNNSF 30771143). The authors would like to acknowledge Professor Wen-lin Wang (Cancer Research Center of Sun Yat-sen University), Dr. Jun-tao Zhou (Department of Anatomy, Sun Yat-sen University) and Dr. Jun-mei Wang.

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Authors and Affiliations

  1. Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, 74 Zhongshan 2nd Road, 510080, Guangzhou, China

    Yan-Qing Zhang, Xiang Zeng, Ying Ding, Yan Li & Yuan-Shan Zeng

  2. Research Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, 510080, Guangzhou, China

    Xiang Zeng & Yuan-Shan Zeng

  3. Institute of Polymer Science, School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou, China

    Liu-Min He

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  1. Yan-Qing Zhang
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  2. Xiang Zeng
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  3. Liu-Min He
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  4. Ying Ding
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  5. Yan Li
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  6. Yuan-Shan Zeng
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Correspondence to Yuan-Shan Zeng.

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12565_2009_56_MOESM1_ESM.tif

S1_Neural cell markers (MAP2) detected in GFP-expressing MSCs by immunofluorescent staining. a TrkC-MSCs; b TrkC-MSCs+NT-3+k252a; c TrkC-MSCs+NT-3. GFP (green) and positively stained cells (red; arrows: yellow cells). Bars a–c 40 μm. d Cells with staining of MAP2 were counted manually. Positively stained cells in the TrkC-MSCs+NT-3 group and TrkC-MSCs+NT-3+k252a group were significantly higher than in the TrkC-MSCs group (*P<0.05). The TrkC-MSCs+NT-3 group also had a significantly higher number of positive cells than the TrkC-MSCs+NT-3+k252a group (△P<0.05) (TIFF 1510 kb)

12565_2009_56_MOESM2_ESM.tif

S2_Neural cell markers (PSD95) detected in GFP-expressing MSCs by immunofluorescent staining.a TrkC-MSCs; b TrkC-MSCs+NT-3+k252a; c TrkC-MSCs+NT-3. GFP (green) and positively stained cells (red; arrows: yellow cells). Bars a–c 40 μm. d Cells with staining of PSD95 were counted manually. Compared with other groups, there were significantly more positively stained cells in the TrkC-MSCs+NT-3 group than in any other group (*P<0.05) (TIFF 1499 kb)

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Zhang, YQ., Zeng, X., He, LM. et al. NT-3 gene modified Schwann cells promote TrkC gene modified mesenchymal stem cells to differentiate into neuron-like cells in vitro. Anat Sci Int 85, 61–67 (2010). https://doi.org/10.1007/s12565-009-0056-8

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  • DOI: https://doi.org/10.1007/s12565-009-0056-8

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