Engineering of adult human neural stem cells differentiation through surface micropatterning

  • Béduer A
  • Vieu C
  • Arnauduc F
 et al. 
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a b s t r a c t Interaction between differentiating neural stem cells and the extracellular environment guides the establishment of cell polarity during nervous system development. Developing neurons read the physical properties of the local substrate in a contact-dependent manner and retrieve essential guidance cues. To restore damage brain area by tissue engineering, the biomaterial scaffold has to mimic this microenvi-ronment to allow organized tissue regeneration. To establish the validity of using microgrooved surfaces in order to simultaneously provide to primary adult human neural stem cells a permissive growth environment and a guide for neurite outgrowth in a pre-established direction, we have studied the long-term culture of adult human neural stem cells from patient biopsies on microgrooved polymers. By exploiting polymer moulding techniques, we engineered non-cytotoxic deep microstructured surfaces of polydimethylsiloxane (PDMS) exhibiting microchannels of various widths. Our results demonstrate that precoated micropatterned PDMS surfaces can serve as effective neurite guidance surfaces for human neural stem cells. Immunocytochemistry analysis show that channel width can impact strongly devel-opment and differentiation. In particular we found an optimal microchannel width, that conciliates a high differentiation rate with a pronounced alignment of neurites along the edges of the micro-channels. The impact of the microstructures on neurite orientation turned out to be strongly influenced by cell density, attesting that cell/surface interactions at the origin of the alignment effect, are in competition with cell/cell interactions tending to promote interconnected networks of cells. Considering all these effects, we have been able to design appropriate structures allowing to obtain neuron devel-opment and differentiation rate comparable to a plane unpatterned surface, with an efficient neurite guidance and a long-term cell viability.

Author-supplied keywords

  • Adult human neural stem cells
  • Bioimplant
  • Cell interactions
  • Microtopography
  • Neurite outgrowth
  • Polymer

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  • Amélie Béduer

  • Christophe Vieu

  • Florent Arnauduc

  • Jean-Christophe Sol

  • Isabelle Loubinoux

  • Laurence Vaysse

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