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Issue 4, 2012
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High-throughput clonal analysis of neural stem cells in microarrayed artificial niches

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Abstract

To better understand the extrinsic signals that control neural stem cell (NSC) fate, here we applied a microwell array platform which allows high-throughput clonal analyses of NSCs, cultured either as neurospheres or as adherent clones, exposed to poly(ethylene glycol) (PEG) hydrogel substrates functionalized with selected signaling molecules. We analyzed by time-lapse microscopy and retrospective immunostaining the role of integrin and Notch ligands, two key NSC niche components, in altering the behavior of several hundred single stem cells isolated from a previously described Hes5::GFP reporter mouse. NSC self-renewal was increased by 1.5-fold upon exposure to covalently tethered Laminin-1 and fibronectin fragment 9–10 (FN9–10), where 60–65% of single cells proliferated extensively and remained Nestin positive. Tethering of the Notch ligand Jagged-1 induced activation of Notch signaling. While Jagged-1 alone increased cell survival and proliferation, no further increase in the clonogenic potential of Hes5::GFP cells was observed upon co-stimulation with Laminin-1 and Jagged-1. We believe that the bioengineering of such in vitro niche analogues is a powerful approach to elucidate single stem cell fate regulation in a well-controlled fashion.

Graphical abstract: High-throughput clonal analysis of neural stem cells in microarrayed artificial niches

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Publication details

The article was received on 14 Jul 2011, accepted on 27 Dec 2011, published on 06 Feb 2012 and first published online on 06 Feb 2012


Article type: Paper
DOI: 10.1039/C2IB00070A
Citation: Integr. Biol., 2012,4, 391-400
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    High-throughput clonal analysis of neural stem cells in microarrayed artificial niches

    M. Roccio, S. Gobaa and M. P. Lutolf, Integr. Biol., 2012, 4, 391
    DOI: 10.1039/C2IB00070A

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