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Use of polyamidoamine dendrimers to engineer BDNF-producing human mesenchymal stem cells

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Abstract

We report the use of polyamidoamine (PAMAM-NH2) dendrimers along with other non-viral vehicles for the in vitro transfection of human bone marrow mesenchymal stem cells (hMSCs) and for engineering MSCs to secrete brain-derived neurotrophic factor (BDNF). Different generations of cationic polyamidoamine dendrimers (generations 3–6) were tested on HEK 293T cells. hMSCs were then transfected with PAMAM-NH2 G4 dendrimers and Lipofectamine 2000, which elicited the expression of GFP reporter in around 6 and 20% of the cells, respectively. Both vehicles were then shown to elicit the expression of BDNF in MSCs from a bicistronic cassette. Non-virally induced neurotrophin expression may be a safe and easy method for adapting autologous stem cells for therapeutic treatment of diseases and neural system injuries.

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References

  1. Ronsyn MW, Daans J, Spaepen G, Chatterjee S et al (2007) Plasmid-based genetic modification of human bone marrow-derived stromal cells:analysis of cell survival and transgene expression after transplantation in rat spinal cord. BMC Biotechnol 7:90–107

    Article  PubMed  Google Scholar 

  2. Ruitenberg MJ, Levison DB, Lee SV, Verhaagen J et al (2005) NT-3 expression from engineered olfactory ensheathing glia promotes spinal sparing and regeneration. Brain 128:839–853

    Article  PubMed  Google Scholar 

  3. Wang LJ, Lu YY, Muramatsu S, Ikeguchi K et al (2002) Neuroprotective effects of glial cell line-derived neurotrophic factor mediated by an adeno-associated virus vector in a transgenic animal model of amyotrophic lateral sclerosis. J Neurosci 22:6920–6928

    CAS  PubMed  Google Scholar 

  4. Weinelt S, Peters S, Bauer P, Mix E et al (2003) Ciliary neurotrophic factor overexpression in neural progenitor cells (ST14A) increases proliferation, metabolic activity, and resistance to stress during differentiation. J Neurosci Res 71:228–236

    Article  CAS  PubMed  Google Scholar 

  5. Ferraro GB, Alabed YZ, Fournier AE (2004) Molecular targets to promote central nervous system regeneration. Curr Neurovasc Res 1:61–75

    Article  CAS  PubMed  Google Scholar 

  6. Reichardt LF (2006) Neurotrophin-regulated signalling pathways. Philos Trans R Soc Lond B Biol Sci 361:1545–1564

    Article  CAS  PubMed  Google Scholar 

  7. Cosgaya JM, Chan JR, Shooter EM (2002) The neurotrophin receptor p75NTR as a positive modulator of myelination. Science 298:1245–1248

    Article  CAS  PubMed  Google Scholar 

  8. Coumans JV, Lin TT, Dai HN, MacArthur L et al (2001) Axonal regeneration and functional recovery after complete spinal cord transection in rats by delayed treatment with transplants and neurotrophins. J Neurosci 21:9334–9344

    CAS  PubMed  Google Scholar 

  9. Heymach JV Jr, Kruttgen A, Suter U, Shooter EM (1996) The regulated secretion and vectorial targeting of neurotrophins in neuroendocrine and epithelial cells. J Biol Chem 271:25430–25437

    Article  CAS  PubMed  Google Scholar 

  10. Kim HK, Lee YS, Sivaprasad U, Malhotra A et al (2006) Muscle-specific microRNA miR-206 promotes muscle differentiation. J Cell Biol 174:677–687

    Article  CAS  PubMed  Google Scholar 

  11. Haensler J, Szoka FC Jr (1993) Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjug Chem 4:372–379

    Article  CAS  PubMed  Google Scholar 

  12. Kukowska-Latallo JF, Bielinska AU, Johnson J, Spindler R et al (1996) Efficient transfer of genetic material into mammalian cells using Starburst polyamidoamine dendrimers. Proc Natl Acad Sci U S A 93:4897–4902

    Article  CAS  PubMed  Google Scholar 

  13. Kuo JH, Lin YL (2007) Remnant cationic dendrimers block RNA migration in electrophoresis after monophasic lysis. J Biotechnol 129:383–390

    Article  CAS  PubMed  Google Scholar 

  14. Svenson S, Tomalia DA (2005) Dendrimers in biomedical applications–reflections on the field. Adv Drug Deliv Rev 57:2106–2129

    Article  CAS  PubMed  Google Scholar 

  15. Santos JL, Oramas E, Pego AP, Granja PL et al (2009) Osteogenic differentiation of mesenchymal stem cells using PAMAM dendrimers as gene delivery vectors. J Control Release 134:141–148

    Article  CAS  PubMed  Google Scholar 

  16. Mowla SJ, Pareek S, Farhadi HF, Petrecca K et al (1999) Differential sorting of nerve growth factor and brain-derived neurotrophic factor in hippocampal neurons. J Neurosci 19:2069–2080

    CAS  PubMed  Google Scholar 

  17. Kurozumi K, Nakamura K, Tamiya T, Kawano Y et al (2004) BDNF gene-modified mesenchymal stem cells promote functional recovery and reduce infarct size in the rat middle cerebral artery occlusion model. Mol Ther 9:189–197

    Article  CAS  PubMed  Google Scholar 

  18. Coppola V, Kucera J, Palko ME, Martinez-De Velasco J et al (2001) Dissection of NT3 functions in vivo by gene replacement strategy. Development 128:4315–4327

    CAS  PubMed  Google Scholar 

  19. Giehl KM, Tetzlaff W (1996) BDNF and NT-3, but not NGF, prevent axotomy-induced death of rat corticospinal neurons in vivo. Eur J Neurosci 8:1167–1175

    Article  CAS  PubMed  Google Scholar 

  20. Ye JH, Houle JD (1997) Treatment of the chronically injured spinal cord with neurotrophic factors can promote axonal regeneration from supraspinal neurons. Exp Neurol 143:70–81

    Article  CAS  PubMed  Google Scholar 

  21. Cao Q, Xu XM, Devries WH, Enzmann GU et al (2005) Functional recovery in traumatic spinal cord injury after transplantation of multineurotrophin-expressing glial-restricted precursor cells. J Neurosci 25:6947–6957

    Article  CAS  PubMed  Google Scholar 

  22. Lu P, Jones LL, Tuszynski MH (2005) BDNF-expressing marrow stromal cells support extensive axonal growth at sites of spinal cord injury. Exp Neurol 191:344–360

    Article  CAS  PubMed  Google Scholar 

  23. Zhang W, Zeng YS, Zhang XB, Wang JM et al (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 

  24. Zhao LX, Zhang J, Cao F, Meng L et al (2004) Modification of the brain-derived neurotrophic factor gene: a portal to transform mesenchymal stem cells into advantageous engineering cells for neuroregeneration and neuroprotection. Exp Neurol 190:396–406

    Article  CAS  PubMed  Google Scholar 

  25. Bertani N, Malatesta P, Volpi G, Sonego P et al (2005) Neurogenic potential of human mesenchymal stem cells revisited:analysis by immunostaining, time-lapse video and microarray. J Cell Sci 118:3925–3936

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Potential conflicts of interest The authors do not have commercial or other associations that might entail a conflict of interest. Sources of financial support This work has been supported by a grant from ERA-NET NAN2007-31198-E.

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Authors and Affiliations

  1. Institute of Genetics and Cytology, National Academy of Sciences of Belarus, Akademicheskaya, 27, 220072, Minsk, Belarus

    Antos Shakhbazau

  2. Institute of Biophysics and Cellular Engineering, National Academy of Sciences of Belarus, Akademicheskaya, 27, 220072, Minsk, Belarus

    Dzmitry Shcharbin

  3. Republic Centre for Haematology and Transfusiology, Minsk, Belarus

    Ihar Seviaryn, Natalya Goncharova, Svetlana Kosmacheva & Mihail Potapnev

  4. Laboratory of Electron Microscopy, University of Lodz, Lodz, Poland

    Barbara Gabara

  5. Department of General Biophysics, University of Lodz, Lodz, Poland

    Maxim Ionov & Maria Bryszewska

Authors
  1. Antos Shakhbazau
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  2. Dzmitry Shcharbin
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  3. Ihar Seviaryn
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  4. Natalya Goncharova
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  5. Svetlana Kosmacheva
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  6. Mihail Potapnev
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  7. Barbara Gabara
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  8. Maxim Ionov
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  9. Maria Bryszewska
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Corresponding authors

Correspondence to Antos Shakhbazau or Dzmitry Shcharbin.

Electronic supplementary material

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11033_2009_9651_MOESM1_ESM.tif

The changes of fluorescence emission intensity of EB complexed with pGFP upon addition of PAMAM G4 dendrimer at different charge ratios. λexc. = 477 nm (TIF 813 kb)

The image of dendriplex “PAMAM G4 – pGFP” at ×80,000. Bar indicates 100 nm (TIF 411 kb)

11033_2009_9651_MOESM3_ESM.tif

Representative flow cytometry analysis of transfection of pGFP by PAMAM G4 dendrimer in cells HEK 293T. Left – control cells, right – G4-transfected cells (TIF 395 kb)

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Shakhbazau, A., Shcharbin, D., Seviaryn, I. et al. Use of polyamidoamine dendrimers to engineer BDNF-producing human mesenchymal stem cells. Mol Biol Rep 37, 2003–2008 (2010). https://doi.org/10.1007/s11033-009-9651-y

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  • DOI: https://doi.org/10.1007/s11033-009-9651-y

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