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Exogenous Expression of Nt-3 and TrkC Genes in Bone Marrow Stromal Cells Elevated the Survival Rate of the Cells in the Course of Neural Differentiation

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Abstract

Bone marrow stromal cells (BMSCs) are attractive cellular sources for cell therapy of many diseases, specifically neurodegenerative ones. The potential capability of BMSCs could be further augmented by enhancing their neuroprotective property, differentiation potential, and survival rate subsequent to transplantation. Therefore, a concurrent upregulation of neurotrophin-3 (NT-3) and its high affinity receptor, tyrosin kinase C (TrkC), was utilized in our study. BMSCs were cotransfected with pDsRed1-N1-NT-3 and pCMX-TrkC plasmids before induction of neural differentiation. pEGFP-N1-transfected BMSCs were also employed as a control. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed for gene expression analysis. Cell viability was evaluated by MTT assay, while apoptosis rate was assessed by flow cytometry after PI and Annexin V staining. NT-3 and TrkC mRNA levels were greatly elevated following cotransfection of cells with pDsRed1-N1-NT-3 and pCMX-TrkC vectors. The expression of neural markers (i.e., NFM, and NeuroD1) was augmented in cotransfected BMSCs, compared to the control ones, after neural induction. At each time point, the viability and apoptosis rates of the cells over-expressing NT-3 and TrkC showed increased and reduced patterns, respectively. Our data demonstrated that NT-3/TrkC-co-transfected BMSCs, compared to those of intact cells, could be more beneficial graft candidates for the upcoming treatment strategies of neurogenic disorders due to their increased viability and expression of neural markers. This may be due to their increased level of neural differentiation potential and/or their enhanced rate of survival and/or their useful capacity to secrete NT-3.

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References

  • Blesch A (2006) Neurotrophic factors in neurodegeneration. Brain Pathol 16:295–303

    Article  CAS  PubMed  Google Scholar 

  • Caplan AI (2007) Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol 213:341–347

    Article  CAS  PubMed  Google Scholar 

  • Chen Q et al (2005) Protective effects of bone marrow stromal cell transplantation in injured rodent brain: synthesis of neurotrophic factors. J Neurosci Res 80:611–619

    Article  CAS  PubMed  Google Scholar 

  • Cheng L, Saphieha P, Kittlerova P, Hauswirth WW, Di Polo AT (2002) TrkB gene transfer protects retinal ganglion cells from axotomy-induced death in vivo. J Neurosci 22:3977–3986

    CAS  PubMed  Google Scholar 

  • Davies AM, Minichiello L, Klein R (1995) Developmental changes in NT-3 signalling via TrkA and TrkB in embryonic neurons. EMBO J 14:4482–4489

    CAS  PubMed Central  PubMed  Google Scholar 

  • Edalat H, Hajebrahimi Z, Movahedin M, Tavallaei M, Amiri S, Mowla SJ (2011) p75NTR suppression in rat bone marrow stromal stem cells significantly reduced their rate of apoptosis during neural differentiation. Neurosci Lett 498:15–19

    Article  CAS  PubMed  Google Scholar 

  • Edalat H, Hajebrahimi Z, Pirhajati V, Movahedin M, Tavallaei M, Soroush MR, Mowla SJ (2013) Transplanting p75-suppressed bone marrow stromal cells promotes functional behavior in a rat model of spinal cord injury. Iran Biomed J 17:140–145

    PubMed Central  PubMed  Google Scholar 

  • Farhadi HF, Mowla SJ, Petrecca K, Morris SJ, Seidah NG, Murphy RA (2000) Neurotrophin-3 sorts to the constitutive secretory pathway of hippocampal neurons and is diverted to the regulated secretory pathway by coexpression with brain-derived neurotrophic factor. J Neurosci 20:4059–4068

    CAS  PubMed  Google Scholar 

  • Gao Z et al (2009) Neurodl is essential for the survival and maturation of adult-born neurons. Nat Neurosci 12:1090–1092

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hajebrahimi Z, Mowla SJ, Movahedin M, Tavallaei M (2008) Gene expression alterations of neurotrophins, their receptors and prohormone convertases in a rat model of spinal cord contusion. Neurosci Lett 441:261–266

    Article  CAS  PubMed  Google Scholar 

  • Hantzopoulos PA, Suri C, Glass DJ, Goldfarb MP, Yancopoulos GD (1994) The low affinity NGF receptor, p75, can collaborate with each of the Trks to potentiate functional responses to the neurotrophins. Neuron 13:187–201

    Article  CAS  PubMed  Google Scholar 

  • Hokari M, Kuroda S, Shichinohe H, Yano S, Hida K, Iwasaki Y (2008) Bone marrow stromal cells protect and repair damaged neurons through multiple mechanisms. J Neurosci Res 86:1024–1035

    Article  CAS  PubMed  Google Scholar 

  • Ip NY et al (1993) Similarities and differences in the way neurotrophins interact with the Trk receptors in neuronal and nonneuronal cells. Neuron 10:137–149

    Article  CAS  PubMed  Google Scholar 

  • Jafarnejad SM, Mowla SJ, Matin MM (2008) Knocking-down the expression of nucleostemin significantly decreases rate of proliferation of rat bone marrow stromal stem cells in an apparently p53-independent manner. Cell Prolif 41:28–35

    Article  CAS  PubMed  Google Scholar 

  • Verge VM, Gratto KA, Karchewski LA, Richardson PM (1996) Neurotrophins and nerve injury in the adult. Philos Trans R Soc Lond B 351:423–430

    Article  CAS  Google Scholar 

  • Yaghoobi MM, Mowla SJ (2006) Differential gene expression pattern of neurotrophins and their receptors during neuronal differentiation of rat bone marrow stromal cells. Neurosci Lett 397:149–154

    Article  CAS  PubMed  Google Scholar 

  • Yaghoobi MM, Mowla SJ, Tiraihi T (2005) Nucleostemin, a coordinator of self-renewal, is expressed in rat marrow stromal cells and turns off after induction of neural differentiation. Neurosci Lett 390:81–86

    Article  CAS  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 

  • Zhang YQ, Zeng X, He LM, Ding Y, Li Y, Zeng YS (2010) 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

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to Dr. Phil Barker for kindly providing us the plasmids. This research was supported by a grant from Basij Elmi Organization (Grant No. 3657).

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Authors and Affiliations

  1. Genetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Houri Edalat & Mahmoud Tavallaei

  2. Department of Physiology, Aerospace Research Institute, Tehran, Iran

    Zahra Hajebrahimi

  3. Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Vahid Pirhajati

  4. Department of Anatomy, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Mansoureh Movahedin

  5. Department of Molecular Genetics, School of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Seyed Javad Mowla

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  1. Houri Edalat
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  2. Zahra Hajebrahimi
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  3. Vahid Pirhajati
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  4. Mahmoud Tavallaei
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  5. Mansoureh Movahedin
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  6. Seyed Javad Mowla
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Correspondence to Seyed Javad Mowla.

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Edalat, H., Hajebrahimi, Z., Pirhajati, V. et al. Exogenous Expression of Nt-3 and TrkC Genes in Bone Marrow Stromal Cells Elevated the Survival Rate of the Cells in the Course of Neural Differentiation. Cell Mol Neurobiol 37, 1187–1194 (2017). https://doi.org/10.1007/s10571-016-0448-y

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  • DOI: https://doi.org/10.1007/s10571-016-0448-y

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