Applied Biochemistry and Biotechnology

, Volume 174, Issue 4, pp 1233–1245 | Cite as

Stimulation of Neural Differentiation in Human Bone Marrow Mesenchymal Stem Cells by Extremely Low-Frequency Electromagnetic Fields Incorporated with MNPs

  • Yun-Kyong Choi
  • Dong Heon Lee
  • Young-Kwon Seo
  • Hyun Jung
  • Jung-Keug Park
  • Hyunjin ChoEmail author


Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) have been investigated as a new cell-therapeutic solution due to their capacity that could differentiate into neural-like cells. Extremely low-frequency electromagnetic fields (ELF-EMFs) therapy has emerged as a novel technique, using mechanical stimulus to differentiate hBM-MSCs and significantly enhance neuronal differentiation to affect cellular and molecular reactions. Magnetic iron oxide (Fe3O4) nanoparticles (MNPs) have recently achieved widespread use for biomedical applications and polyethylene glycol (PEG)-labeled nanoparticles are used to increase their circulation time, aqueous solubility, biocompatibility, and nonspecific cellular uptake as well as to decrease immunogenicity. Many studies have used MNP-labeled cells for differentiation, but there have been no reports of MNP-labeled neural differentiation combined with EMFs. In this study, synthesized PEG-phospholipid encapsulated magnetite (Fe3O4) nanoparticles are used on hBM-MSCs to improve their intracellular uptake. The PEGylated nanoparticles were exposed to the cells under 50 Hz of EMFs to improve neural differentiation. First, we measured cell viability and intracellular iron content in hBM-MSCs after treatment with MNPs. Analysis was conducted by RT-PCR, and immunohistological analysis using neural cell type-specific genes and antibodies after exposure to 50 Hz electromagnetic fields. These results suggest that electromagnetic fields enhance neural differentiation in hBM-MSCs incorporated with MNPs and would be an effective method for differentiating neural cells.


Extremely low-frequency electromagnetic fields Bone marrow-derived mesenchymal stem cell Magnetic nanoparticle Neural differentiation 



This research was supported by the Pioneer Research Center Program through the National Research Foundation of Korea funded by the Ministry of Science, ICT & Future Planning (grant number 2009-0082941).


  1. 1.
    Barry, F. P., & Murphy, J. M. (2004). Mesenchymal stem cells: clinical applications and biological characterization. International Journal of Biochemistry & Cell Biology, 36(4), 568–584.CrossRefGoogle Scholar
  2. 2.
    Salem, H. K., & Thiemermann, C. (2010). Mesenchymal stromal cells: current understanding and clinical status. Stem Cells, 28(3), 585–596.Google Scholar
  3. 3.
    Izadpanah, R., Trygg, C., Patel, B., et al. (2006). Biologic properties of mesenchymal stem cells derived from bone marrow and adipose tissue. Journal of Cellular Biochemistry, 99(5), 1285–1297.CrossRefGoogle Scholar
  4. 4.
    Pountos, I., & Giannoudis, P. V. (2005). Biology of mesenchymal stem cells. Injury, 36, S8–S12.CrossRefGoogle Scholar
  5. 5.
    Pittenger, M. F., Mackay, A. M., Beck, S. C., et al. (1999). Multilineage potential of adult human mesenchymal stem cells. Science, 284(5411), 143–147.CrossRefGoogle Scholar
  6. 6.
    Prockop, D. J. (1997). Marrow stromal cells as stem cells for nonhematopoietic tissues. Science, 276(5309), 71–74.CrossRefGoogle Scholar
  7. 7.
    Woodbury, D., Schwarz, E. J., Prockop, D. J., et al. (2000). Adult rat and human bone marrow stromal cells differentiate into neurons. Journal of Neuroscience Research, 61(4), 364–370.CrossRefGoogle Scholar
  8. 8.
    Sanchez-Ramos, J., Song, S., Cardozo-Pelaez, F., et al. (2000). Adult bone marrow stromal cells differentiate into neural cells in vitro. Experimental Neurology, 164(2), 247–256.CrossRefGoogle Scholar
  9. 9.
    Lacy-Hulbert, A., Metcalfe, J., & Hesketh, R. (1998). Biological responses to electromagnetic fields. FASEB Journal, 12(6), 395–420.Google Scholar
  10. 10.
    Sun, S., Liu, Y., Lipsky, S., et al. (2007). Physical manipulation of calcium oscillations facilitates osteo differentiation of human mesenchymal stem cells. FASEB Journal, 21(7), 1472–1480.CrossRefGoogle Scholar
  11. 11.
    Sert, C., Mustafa, D., Duz, M. Z., et al. (2002). The preventive effect on bone loss of 50-Hz, 1-mT electromagnetic field in ovariectomized rats. Journal of Bone and Mineral Metabolism, 20(6), 345–349.CrossRefGoogle Scholar
  12. 12.
    McLeod, K. J., & Rubin, C. T. (1992). The effect of low-frequency electrical fields on osteogenesis. Journal of Bone and Joint Surgery (American), 74(6), 920–929.CrossRefGoogle Scholar
  13. 13.
    Piacentini, R., Ripoli, C., Mezzogori, D., et al. (2008). Extremely low frequency electromagnetic fields promote in vitro neurogenesis via upregulation of Ca(v)1-channel activity. Journal of Cellular Physiology, 215(1), 129–139.CrossRefGoogle Scholar
  14. 14.
    Cuccurazzu, B., Leone, L., Podda, M. V., et al. (2010). Exposure to extremely low-frequency (50 Hz) electromagnetic fields enhances adult hippocampal neurogenesis in C57BL/6 mice. Experimental Neurology, 226(1), 173–182.CrossRefGoogle Scholar
  15. 15.
    Cho, H., Seo, Y. K., Yoon, H. H., et al. (2012). Neural stimulation on human bone marrow-derived mesenchymal stem cells by extremely low frequency electromagnetic fields. Biotechnology Progress, 28(5), 1329–1335.CrossRefGoogle Scholar
  16. 16.
    Lattuada, M., & Hatton, T. A. (2007). Functionalization of monodisperse magnetic nanoparticles. Langmuir, 23(4), 2158–2168.CrossRefGoogle Scholar
  17. 17.
    Pankhurst, Q. A., Connolly, J., Jones, S. K., et al. (2003). Applications of magnetic nanoparticles in biomedicine. Journal of Physics D: Applied Physics, 36, R167.CrossRefGoogle Scholar
  18. 18.
    Xie, J., Xu, C., Kohler, N., et al. (2007). Controlled PEGylation of monodisperse Fe3O4 Nanoparticles for reduced non‐specific uptake by macrophage cells. Advanced Materials, 19, 2163–3166.CrossRefGoogle Scholar
  19. 19.
    Greenwald, R. B., Choe, Y. H., McGuire, J., et al. (2003). Effective drug delivery by PEGylated drug conjugates. Advanced Drug Delivery Reviews, 55(2), 217–250.CrossRefGoogle Scholar
  20. 20.
    Schäfer, R., Kehlbach, R., Müller, M., et al. (2009). Labeling of human mesenchymal stromal cells with superparamagnetic iron oxide leads to a decrease in migration capacity and colony formation ability. Cytotherapy, 11(1), 68–78.CrossRefGoogle Scholar
  21. 21.
    Schäfer, R., Bantleon, R., Kehlbach, R., et al. (2010). Functional investigations on human mesenchymal stem cells exposed to magnetic fields and labeled with clinically approved iron nanoparticles. BMC Cell Biology, 11, 22.CrossRefGoogle Scholar
  22. 22.
    Shimizu, K., Ito, A., Yoshida, T., et al. (2007). Bone tissue engineering with human mesenchymal stem cell sheets constructed using magnetite nanoparticles and magnetic force. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 82(2), 471–480.CrossRefGoogle Scholar
  23. 23.
    Qian, D. X., Zhang, H. T., Ma, X., et al. (2010). Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells. Neurochemical Research, 35(4), 572–579.CrossRefGoogle Scholar
  24. 24.
    Cho, H., Choi, Y. K., Lee, D. H., et al. (2013). Effects of magnetic nanoparticle-incorporated human bone marrow-derived mesenchymal stem cells exposed to pulsed electromagnetic fields on injured rat spinal cord. Biotechnology and Applied Biochemistry, 60(6), 596–602.CrossRefGoogle Scholar
  25. 25.
    Rivers, F. J., Couillard-Despres, S., Pedre, X., et al. (2006). Mesenchymal stem cells instruct oligodendrogenic fate decision on adult neural stem cells. Stem Cells, 24(10), 2209–2219.CrossRefGoogle Scholar
  26. 26.
    Bahat-Stroomza, B., Barhum, Y., Levy, Y. S., et al. (2009). Induction of adult human bone marrow mesenchymal stromal cells into functional astrocyte-like cells: potential for restorative treatment in parkinson’s disease. Journal of Molecular Neuroscience, 39(1–2), 199–210.CrossRefGoogle Scholar
  27. 27.
    McCaig, C. D., Sangster, L., & Stewart, R. (2000). Neurotrophins enhance electric field-directed growth cone guidance and directed nerve branching. Developmental Dynamics, 217(3), 299–308.CrossRefGoogle Scholar
  28. 28.
    Sisken, B. F., Walker, J., & Orgel, M. (1993). Prospects on clinical applications of electrical stimulation for nerve regeneration. Journal of Cellular Biochemistry, 51(4), 404–409.CrossRefGoogle Scholar
  29. 29.
    McCaig, C. D. (1986). Electric fields, contact guidance and the direction of nerve growth. Journal of Embryology & Experimental Morphology, 94, 245–255.Google Scholar
  30. 30.
    Kunzmann, A., Andersson, B., Thurnherr, T., et al. (2011). Toxicology of engineered nanomaterials: focus on biocompatibility, biodistribution and biodegradation. Biochimica et Biophysica Acta, 1810(3), 361–373.CrossRefGoogle Scholar
  31. 31.
    Zhang, Y., Kohler, N., & Zhang, M. (2002). Surface modification of superparamagnetic magnetite nanoparticles and their intracellular uptake. Biomaterials, 23(7), 1553–1561.CrossRefGoogle Scholar
  32. 32.
    Riggio, C., Calatayud, M. P., Hoskins, C., et al. (2012). Poly-l-lysine-coated magnetic nanoparticles as intracellular acutators for neural guidance. International Journal of Nanomedicine, 7, 3155–3166.Google Scholar
  33. 33.
    Safford, K. M., Hicok, K. C., Safford, S. D., et al. (2002). Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochemical and Biophysical Research Communications, 294(2), 371–379.CrossRefGoogle Scholar
  34. 34.
    Mitchell, K. E., Weiss, M. L., Mitchell, B. M., et al. (2003). Matrix cells from Wharton’s jelly form neurons and glia. Stem Cells, 21(1), 50–60.CrossRefGoogle Scholar
  35. 35.
    Tseng, P. Y., Chen, C. J., Sheu, C. C., et al. (2007). Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells. Journal of Veterinary Medical Science, 69(2), 95–102.CrossRefGoogle Scholar
  36. 36.
    Ge, D., Song, K., Guan, S., et al. (2013). Culture and differentiation of rat neural stem/progenitor cells in a three-dimensional collagen scaffold. Applied Biochemistry and Biotechnology, 170(2), 406–419.CrossRefGoogle Scholar
  37. 37.
    Zhu, X. L., Eibl, O., Scheideler, L., et al. (2006). Characterization of nano hydroxyapatite/collagen surfaces and cellular behaviors. Journal of Biomedical Materials Research, Part A, 79A, 114–127.CrossRefGoogle Scholar
  38. 38.
    Bock, N., Riminucci, A., Dionigi, C., et al. (2010). A novel route in bone tissue engineering: magnetic biomimetic scaffolds. Acta Biomaterialia, 6(3), 786–796.CrossRefGoogle Scholar
  39. 39.
    Badaracco, M. E., Siri, M. V., & Pasquini, J. M. (2010). Oligodendrogenesis: the role of iron. Biofactors, 36(2), 98–102.Google Scholar
  40. 40.
    Todorich, B., Pasquini, J. M., Garcia, C. I., et al. (2009). Oligodendrocytes and myelination: the role of iron. Glia, 57(5), 467–478.CrossRefGoogle Scholar
  41. 41.
    Kim, J. A., Lee, N., Kim, B. H., et al. (2011). Enhancement of neurite outgrowth in PC12 cells by iron oxide nanoparticles. Biomaterials, 32(11), 2871–2877.CrossRefGoogle Scholar
  42. 42.
    Curran, J. M., Pu, F., Chen, R., et al. (2011). The use of dynamic surface chemistries to control msc isolation and function. Biomaterials, 32(21), 4753–4760.CrossRefGoogle Scholar
  43. 43.
    Bito, H., & Takemoto-Kimura, S. (2003). Ca(2+)/CREB/CBP-dependent gene regulation: a shared mechanism critical in long-term synaptic plasticity and neuronal survival. Cell Calcium, 34(4–5), 425–430.CrossRefGoogle Scholar
  44. 44.
    Wang, J., Weaver, I. C., Gauthier-Fisher, A., et al. (2010). CBP histone acetyltransferase activity regulates embryonic neural differentiation in the normal and Rubinstein–Taybi syndrome brain. Developmental Cell, 18(1), 114–125.CrossRefGoogle Scholar
  45. 45.
    Nakagawa, S., Kim, J. E., Lee, R., et al. (2002). Regulation of neurogenesis in adult mouse hippocampus by cAMP and the cAMP response element-binding protein. Journal of Neuroscience, 22(9), 3673–3682.Google Scholar
  46. 46.
    Leone, L., Fusco, S., Mastrodonato, A., et al. (2014). Epigenic modulation of adult hippocampal neurogenesis by extremely low-frequency electromagnetic fields. Molecular Neurobiology, 49(3), 1472–1486.CrossRefGoogle Scholar
  47. 47.
    Podda, M. V., Leone, L., Barbati, S. A., et al. (2014). Extremely low-frequency electromagnetic fields enhance the survival of newborn neurons in the mouse hippocampus. European Journal of Neuroscience, 39(6), 893–903.CrossRefGoogle Scholar
  48. 48.
    Park, J. E., Seo, Y. K., Yoon, H. H., et al. (2013). Electromagnetic fields induce neural differentiation of human bone marrow derived mesenchymal stem cells via ROS mediated EGRF activation. Neurochemistry International, 62(4), 418–424.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Yun-Kyong Choi
    • 1
  • Dong Heon Lee
    • 2
  • Young-Kwon Seo
    • 1
  • Hyun Jung
    • 2
  • Jung-Keug Park
    • 1
    • 3
  • Hyunjin Cho
    • 3
    Email author
  1. 1.Department of Medical BiotechnologyDongguk UniversitySeoulSouth Korea
  2. 2.Advanced Functional Nanohybrid Material Laboratory, Department of ChemistryDongguk UniversitySeoulSouth Korea
  3. 3.Dongguk University Research Institute of BiotechnologySeoulSouth Korea

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