Skip to main content
Log in

Networked neuro-spheres formed by topological attractants for engineering of 3-dimensional nervous system

  • Original Article
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

The nervous systems including central and peripheral nervous system have an important role in transmitting signals from brain to organs and tissues. Due to such critical function of nervous system, considerable effort has been tried to establish in vitro nervous model. In this paper, the neuro-spheres networked by the nerve-like structure were created using concave well arrays connected by the hemicylindrical channels. The concave microwells and the hemicylindrical channels were fabricated using the surface tension of viscose liquid PDMS prepolymer for the creation of neuro-spheres-networking (NSN). To investigate the topological effect of the concave-well-hemicylindrical-channel-networking (CWHCN), comparative experiments were conducted on a conventional cylindrical-wells-rectangular-channel-networking (CWRCN). Neuro-progenitor cells from the rat were seeded on the concave well arrays connected by the hemicylindrical channels and cultured for 10 days. Small neuro-pehroids consisting of neurons and glia cells were autonomously formed in the concave microwell arrays. These neuro-spheres were networked by the nerve-like structures formed along the CWHCN. In order to confirm the interconnection of the neurites in the NSN, a calcium imaging experiment was performed to measure the calcium flux along the nerve-like networking. To demonstrate further use of the networked neuro-networking for brain regeneration, we transferred the NSN onto hydrogel maintaining approximately 90% viability, and this model is expected to be used for the regeneration of a damaged brain.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. M Kato-Negishi, Y Tsuda, H Onoe, et al., A neurospheroid network-stamping method for neural transplantation to the brain, Biomaterial, 31, 34 (2010).

    Article  Google Scholar 

  2. YJ Choi, J Park, and S-H Lee, Size-controllable networked neurospheres as a 3D neuronal tissue model for Alzheimer’s disease studies, Biomaterials, 34, 12 (2013).

    Google Scholar 

  3. M Kato-Negishi, Y Morimoto, H Onoe, et al., Millimeter-Sized Neural Building Blocks for 3D Heterogeneous Neural Network Assembly, Advanced healthcare materials, 2, 12 (2013).

    Article  Google Scholar 

  4. C Patrick, Neural tissue engineering, Annals of Biomedical Engineering, 25, 1 (1997).

    Article  Google Scholar 

  5. CE Schmidt and JB Leach, Neural tissue engineering: strategies for repair and regeneration, Annual review of biomedical engineering, 5, 1 (2003).

    Article  Google Scholar 

  6. J Park, E Lim, S Back, et al., Nerve regeneration following spinal cord injury using matrix metalloproteinase-sensitive, hyaluronic acid-based biomimetic hydrogel scaffold containing brain-derived neurotrophic factor, Journal of Biomedical Materials Research Part A, 93, 3 (2009).

    Google Scholar 

  7. SK Kwon and CG Cho, Regeneration of Facial Nerve using Mesenchymal Stem Cells in Facial Nerve Palsy Animal Model, TISSUE ENGINEERING AND REGENERATIVE MEDICINE, 6, 1 (2009).

    Google Scholar 

  8. GRD Evans, Peripheral nerve injury: a review and approach to tissue engineered constructs, The Anatomical Record, 263, 4 (2001).

    Article  Google Scholar 

  9. PG Gross, EP Kartalov, A Scherer, et al., Applications of microfluidics for neuronal studies, Journal of the neurological sciences, 252, 2 (2007).

    Article  Google Scholar 

  10. Y-J Gao and R-R Ji, Chemokines, neuronal-glial interactions, and central processing of neuropathic pain, Pharmacology & therapeutics, 126, 1 (2010).

    Article  Google Scholar 

  11. RJ Miller and PB Tran, Chemokinetics, Neuron, 47, 5 (2005).

    Article  Google Scholar 

  12. SM Kim, SH Kim, CM Kim, et al., Effect of PLGA/DBP scaffolds seeded OECs and SCs on the proliferation and differentiation of BMSCs, Tissue Eng Regen Med, 4, 4 (2007).

    Google Scholar 

  13. JB Recknor, DS Sakaguchi, and SK Mallapragada, Directed growth and selective differentiation of neural progenitor cells on micropatterned polymer substrates, Biomaterials, 27, 22 (2006).

    Article  Google Scholar 

  14. F Yang, R Murugan, S Wang, et al., Electrospinning of nano/micro scale poly (L-lactic acid) aligned fibers and their potential in neural tissue engineering, Biomaterials, 26, 15 (2005).

    Google Scholar 

  15. E Kang, GS Jeong, YY Choi, et al., Digitally tunable physicochemical coding of material composition and topography in continuous microfibres, Nature materials, 10, 11 (2011).

    Article  Google Scholar 

  16. E Kang, YY Choi, S-K Chae, et al., Microfluidic Spinning of Flat Alginate Fibers with Grooves for Cell-aligning Scaffolds, Advanced Materials, 24, 31 (2012).

    Google Scholar 

  17. SB Jun, MR Hynd, N Dowell-Mesfin, et al., Low-density neuronal networks cultured using patterned poly-l-lysine on microelectrode arrays, Journal of neuroscience methods, 160, 2 (2007).

    Article  Google Scholar 

  18. W Jang, S Kim, I Lee, et al., Neurogenesis of bone marrow stromal cell using controlled release of butylated hydroxyanisole from PLGA films. Tissue Eng Regen Med, 2, 2 (2005).

    Google Scholar 

  19. HG Sundararaghavan, GA Monteiro, BL Firestein, et al., Neurite growth in 3D collagen gels with gradients of mechanical properties, Biotechnology and bioengineering, 102, 2 (2009).

    Article  Google Scholar 

  20. TC Holmes, S de Lacalle, X Su, et al., Extensive neurite outgrowth and active synapse formation on self-assembling peptide scaffolds, Proceedings of the National Academy of Sciences, 97, 12 (2000).

    Article  Google Scholar 

  21. LF Reichardt and KJ Tomaselli, Extracellular matrix molecules and their receptors: functions in neural development, Annual review of neuroscience, 14, (1991).

    Google Scholar 

  22. TA Kapur and MS Shoichet, Immobilized concentration gradients of nerve growth factor guide neurite outgrowth, Journal of Biomedical Materials Research Part A, 68, 2 (2004).

    Google Scholar 

  23. CR Kothapalli, E van Veen, S de Valence, et al., A highthroughput microfluidic assay to study neurite response to growth factor gradients, Lab on a chip, 11, 3 (2011).

    Article  Google Scholar 

  24. SW Rhee, AM Taylor, CH Tu, et al., Patterned cell culture inside microfluidic devices, Lab on a chip, 5, 1 (2005).

    Article  Google Scholar 

  25. NS Baek, YH Kim, YH Han, et al., Facile photopatterning of polyfluorene for patterned neuronal networks, Soft Matter, 7, 21 (2011).

    Article  Google Scholar 

  26. R Segev, M Benveniste, E Hulata, et al., Long term behavior of lithographically prepared in vitro neuronal networks, Physical review letters, 88, 11 (2002).

    Article  Google Scholar 

  27. R Segev, M Benveniste, Y Shapira, et al., Formation of electrically active clusterized neural networks, Physical review letters, 90, 16 (2003).

    Article  Google Scholar 

  28. T Gabay, E Jakobs, E Ben-Jacob, et al., Engineered selforganization of neural networks using carbon nanotube clusters, Physica A: Statistical Mechanics and its Applications, 350, 2 (2005).

    Article  Google Scholar 

  29. GS Jeong, Y Jun, JH Song, et al., Meniscus induced self organization of multiple deep concave wells in a microchannel for embryoid bodies generation, Lab on a chip, 12, 1 (2012).

    Article  Google Scholar 

  30. GS Jeong, JH Song, AR Kang, et al., Surface Tension-mediated, Concave-microwell Arrays for Large-scale, Simultaneous Production of Homogeneously Sized Embryoid Bodies, Advanced healthcare materials, 2, 1 (2013).

    Article  Google Scholar 

  31. SF Wong, DY No, YY Choi, et al., Concave microwell based size-controllable hepatosphere as a three-dimensional liver tissue model, Biomaterials, 32, 32 (2011).

    Google Scholar 

  32. S-A Lee, YY Choi, D Park, et al., Functional 3D human primary hepatocyte spheroids made by co-culturing hepatocytes from partial hepatectomy specimens and human adiposederived stem cells, PloS one, 7, 12 (2012).

    Google Scholar 

  33. Y Jun, AR Kang, JS Lee, et al., 3D co-culturing model of primary pancreatic islets and hepatocytes in hybrid spheroid to overcome pancreatic cell shortage, Biomaterials, 34, 15 (2013).

    Google Scholar 

  34. F Bahner, EK Weiss, G Birke, et al., Cellular correlate of assembly formation in oscillating hippocampal networks in vitro, Proceedings of the National Academy of Sciences, 108, 35 (2011).

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Gi Seok Jeong.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jeong, G.S. Networked neuro-spheres formed by topological attractants for engineering of 3-dimensional nervous system. Tissue Eng Regen Med 11, 297–303 (2014). https://doi.org/10.1007/s13770-014-4047-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13770-014-4047-z

Key words

Navigation