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Neural Stem Cell Fate Control on Micropatterned Substrates

  • Leonora BuzanskaEmail author
  • Marzena Zychowicz
  • Ana Ruiz
  • François Rossi
Protocol
Part of the Neuromethods book series (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.

Key words

Neural stem cells Piezoelectric microspotting Microcontact printing Bioactive domains 

Notes

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.”

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Leonora Buzanska
    • 1
    Email author
  • Marzena Zychowicz
    • 2
  • Ana Ruiz
    • 3
  • François Rossi
    • 3
  1. 1.Mossakowski Medical Research Centre, Stem Cell Bioengineering UnitPolish Academy of SciencesWarsawPoland
  2. 2.Stem Cell Bioengineering Unit, Mosakowski Medical Research CentrePolish Academy of SciencesWarsawPoland
  3. 3.European Commission, Joint Research CentreInstitute for Health and Consumer ProtectionIspraItaly

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