Skip to main content
SpringerLink
Log in
Book cover

Stem Cell Technologies in Neuroscience pp 19–44Cite as

Neural Stem Cell Fate Control on Micropatterned Substrates

  • Protocol
  • First Online:

Part of the book series: Neuromethods ((NM,volume 126))

Abstract

Neural stem cell fate decisions are dependent upon signals coming from microenvironment composed of extracellular matrix proteins, soluble factors and specificity of cell–cell contacts. We have developed in vitro systems to trace developmental processes of neural stem cells and to control their fate commitment. Applied technologies include nano/micro-fabrication techniques, like microcontact printing and piezoelectric microspotting of biomolecules on plasma deposited cell repellent surface. Designed bioactive domains with controlled content and geometry served as a template to immobilize neural stem cells to the surface and direct their differentiation. Migration and axon-like outgrowth have been successfully guided by means of interconnected squares configuration. Receptor mediated versus electrostatic interactions on the cell membrane–surface interface were crucial to keep the cells either in neurally committed or in non-differentiated stages by fibronectin or poly-L-lysine pattern, respectively. Single cell versus multicellular positioning further promoted stem cells non-differentiated stage. Activation of intracellular pathways by signaling molecules (Wnt, CNTF, Jagged, Notch and DKK-1) microspotted with fibronectin directed differentiation into astrocytic and neuronal lineages, as revealed by immunocytochemical and molecular analysis. Our results proved that neural stem cell fate decisions can be influenced by manipulating the composition and architecture of the 2D bioactive domains reflecting their natural niche microenvironment.

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

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   119.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Lutolf MP, Gilbert PM, Blau HM (2009) Designing materials to direct stem-cell fate. Nature 462(7272):433–441

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Stabenfeldt SE, Munglani G, García AJ, LaPlaca MC (2010) Biomimetic microenvironment modulates neural stem cell survival, migration, and differentiation. Tissue Eng Part A 16(12):3747–3758

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Riquelme PA, Drapeau E, Doetsch F (2008) Brain micro-ecologies: neural stem cell niches in the adult mammalian brain. Philos Trans R Soc Lond B Biol Sci 363(1489):123–137

    Article  PubMed  Google Scholar 

  4. Lee MR, Kwon KW, Jung H, Kim HN, Suh KY, Kim K, Kim KS (2010) Direct differentiation of human embryonic stem cells into selective neurons on nanoscale ridge/groove pattern arrays. Biomaterials 31(15):4360–4366

    Article  CAS  PubMed  Google Scholar 

  5. Vecino E, Heller JP, Veiga-Crespo P, Martin KR, Fawcett JW (2015) Influence of extracellular matrix components on the expression of integrins and regeneration of adult retinal ganglion cells. PLoS One 10(5):e0125250

    Article  PubMed  PubMed Central  Google Scholar 

  6. Kazanis I, Lathia JD, Vadakkan TJ, Raborn E, Wan R, Mughal MR, Eckley DM, Sasaki T, Patton B, Mattson MP, Hirschi KK, Dickinson ME, ffrench-Constant C (2010) Quiescence and activation of stem and precursor cell populations in the subependymal zone of the mammalian brain are associated with distinct cellular and extracellular matrix signals. J Neurosci 21:9771–9781

    Article  Google Scholar 

  7. Ruiz A, Buzanska L, Gilliland D, Rauscher H, Sirghi L, Sobanski T, Zychowicz M, Ceriotti L, Bretagnol F, Coecke S, Colpo P, Rossi F (2008) Micro-stamped surfaces for the patterned growth of neural stem cells. Biomaterials 29(36):4766–4774

    Article  CAS  PubMed  Google Scholar 

  8. Bużańska L, Zychowicz M, Ruiz A, Ceriotti L, Coecke S, Rausher H, Sobanski T, Wheland M, Domanska-Janik K, Colpo P, Rossi F (2010) Neural stem cells from human cord blood on bioengineered surfaces—novel approach to multiparameter bio-tests. Toxicology 270:35–42

    Article  PubMed  Google Scholar 

  9. Zychowicz M, Mehn D, Ana Ruiz A, Colpo P, Francois Rossi F, Frontczak-Baniewicz M, Domanska-Janik K, Buzanska L (2011) Proliferation capacity of cord blood derived neural stem cell line on different micro-scale biofunctional domains. Acta Neurobiol Exp (Wars) 71(1):12–23

    Google Scholar 

  10. Béduer A, Vieu C, Arnauduc F, Sol JC, Loubinoux I, Vaysse L (2012) Engineering of adult human neural stem cells differentiation through surface micropatterning. Biomaterials 33(2):504–514

    Article  PubMed  Google Scholar 

  11. Joo S, Kim JY, Lee E, Hong N, Sun W, Nam Y (2015) Effects of ECM protein micropatterns on the migration and differentiation of adult neural stem cells. Sci Rep 5:13043

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Saha K, Pollock JF, Schaffer DV, Healy KE (2007) Designing synthetic materials to control stem cell phenotype. Curr Opin Chem Biol 11(4):381–387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Bakhru S, Nain AS, Highley C, Wang J, Campbell P, Amon C, Zappe S (2011) Direct and cell signaling-based, geometry-induced neuronal differentiation of neural stem cells. Integr Biol (Camb) 3(12):1207–1214

    Article  CAS  Google Scholar 

  14. Dertinger SK, Jiang X, Li Z, Murthy VN, Whitesides GM (2002) Gradients of substrate-bound laminin orient axonal specification of neurons. Proc Natl Acad Sci USA 99(20):12542–12547

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Millet LJ, Stewart ME, Nuzzo RG, Gillette MU (2010) Guiding neuron development with planar surface gradients of substrate cues deposited using microfluidic devices. Lab Chip 10(12):1525–1535

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Dalby MJ, Andar A, Nag A, Affrossman S, Tare R, McFarlane S, Oreffo RO (2008) Genomic expression of mesenchymal stem cells to altered nanoscale topographies. J R Soc Interface 5(26):1055–1065

    Article  PubMed  PubMed Central  Google Scholar 

  17. Kim WR, Jang MJ, Joo S, Sun W, Nam Y (2014) Surface-printed microdot array chips for the quantification of axonal collateral branching of a single neuron in vitro. Lab Chip 14(4):799–805

    Article  CAS  PubMed  Google Scholar 

  18. Zychowicz M, Mehn D, Ruiz A, Frontczak-Baniewicz M, Rossi F, Buzanska L (2012) Patterning of human cord blood-derived stem cells on single cell posts and lines: implications for neural commitment. Acta Neurobiol Exp (Wars) 72(4):325–336

    Google Scholar 

  19. Soen Y, Mori A, Palmer TD, Brown PO (2006) Exploring the regulation of human neural precursor cell differentiation using arrays of signaling microenvironments. Mol Syst Biol 2:37

    Article  PubMed  PubMed Central  Google Scholar 

  20. Bużańska L, Jurga M, Stachowiak EK, Stachowiak MK, Domanska-Janik K (2006) Neural stem-like cell line derived from a nonhematopoietic population of human umbilical cord blood. Stem Cells Dev 15:391–406

    Article  PubMed  Google Scholar 

  21. Ruiz A, Zychowicz M, Ceriotti L, Mehn D, Sirghi L, Rauscher H, Mannelli I, Colpo P, Buzanska L, Rossi F (2013) Microcontact printing and microspotting as methods for direct protein patterning on plasma deposited polyethylene oxide: application to stem cell patterning. Biomed Microdevices 15(3):495–507

    Article  CAS  PubMed  Google Scholar 

  22. Brétagnol F, Lejeune M, Papadopoulou-Bouraoui A, Hasiwa M, Rauscher H, Ceccone G, Colpo P, Rossi F (2006) Fouling and non-fouling surfaces produced by plasma polymerization of ethylene oxide monomer. Acta Biomater 2(2):165–172

    Article  PubMed  Google Scholar 

  23. Bretagnol F, Ceriotti L, Lejeune M, Papadopoulou A, Hasiwa M, Gilliland D, Ceccone G, Colpo P, Rossi F (2006) Functional micropatterned surfaces by combination of plasma polymerization and lift-off processes. Plasma Process Polym 3(1):30–38

    Article  CAS  Google Scholar 

  24. Ruiz A, Ceriotti L, Buzanska L, Hasiwa M, Bretagnol F, Ceccone G, Gilliland D, Rauscher H, Coecke S, Colpo P, Rossi F (2007) Controlled micropatterning of biomolecules for cell culturing. Microelectron Eng 84:1733–1736

    Article  CAS  Google Scholar 

  25. Ceriotti L, Buzanska L, Rausche rH, Mannelli I, Sirghi L, Gilliland D, Hasiwa M, Bretagnol F, Zychowicz M, Ruiz A, Bremer S, Coecke S, Colpo P, Rossi F (2009) Fabrication and characterization of protein arrays for stem cell patterning. Soft Matter 5:1406–1416

    Article  CAS  Google Scholar 

  26. Buzanska L, Ruiz A, Zychowicz M, Rausher H, Ceriotti L, Rossi F, Colpo P, Domanska-Janik K, Coecke S (2009) Patterned growth and differentiation of human cord blood-derived neural stem cells on bio-functionalized surfaces. Acta Neurobiol Exp (Wars) 69:1–14

    Google Scholar 

  27. Guemouri L, Ogier J, Zekhnini Z, Ramsden JJ (2000) The architecture of fibronectin at surfaces. J Chem Phys 113:8183

    Article  CAS  Google Scholar 

  28. Ruiz A, Zychowicz M, Buzanska L, Mehn D, Mills CA, Martinez E, Coecke S, Samitier J, Colpo P, Rossi F (2009) Single stem cell positioning on polylysine and fibronectin microarrays. Micro Nanosyst 1:50–56

    Article  CAS  Google Scholar 

  29. Margadant C, van Opstal A, Boonstra J (2007) Focal adhesion signaling and actin stress fibers are dispensable for progression through the ongoing cell cycle. J Cell Sci 120:66–76

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to Dr Laura Ceriotti for the participation in the microspotting project. We also thank Sabrina Gioria, Patricia Lisboa, G. Ceccone, and Hubert Rauscher for their scientific and technical assistance. This work was supported by statutory founds to MMRC and by Polish Ministry of Scientific Research and Higher Education grant No: 2211/B/P01/2010/38 and European Commission Joint Research Centre Actions “NanoBiotechnology for Health” and “Validation for Consumer Products.”

Author information

Authors and Affiliations

  1. Mossakowski Medical Research Centre, Stem Cell Bioengineering Unit, Polish Academy of Sciences, 5 Pawinskiego St., 02-106, Warsaw, Poland

    Leonora Buzanska

  2. Stem Cell Bioengineering Unit, Mosakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland

    Marzena Zychowicz

  3. European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Ispra, Italy

    Ana Ruiz & François Rossi

Authors
  1. Leonora Buzanska
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marzena Zychowicz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ana Ruiz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. François Rossi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Leonora Buzanska .

Editor information

Editors and Affiliations

  1. Department of Pediatric Surgery Children’s Program in Regenerative Medicine, University of Texas Health Science Center at Houston McGovern Medical School, Houston, Texas, USA

    Amit K. Srivastava

  2. Center for Stem Cells and Regenerative Medicine, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, USA

    Evan Y. Snyder

  3. Departments of PM&R and Neurosurgery Harvard Medical School, Spaulding Rehabilitation Network and Brigham & Women’s Hospital, Boston, Massachusetts, USA

    Yang D. Teng

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Buzanska, L., Zychowicz, M., Ruiz, A., Rossi, F. (2017). Neural Stem Cell Fate Control on Micropatterned Substrates. In: Srivastava, A., Snyder, E., Teng, Y. (eds) Stem Cell Technologies in Neuroscience. Neuromethods, vol 126. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7024-7_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7024-7_2

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7022-3

  • Online ISBN: 978-1-4939-7024-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation