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Enhancing the reliability and throughput of neurosphere culture on hydrogel microwell arrays.

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  • 2008-08-02
Published in:
  • Stem cells (Dayton, Ohio). - 2008
Animals
Biological Assay
Cell Culture Techniques
Cell Lineage
Cell Movement
Cell Proliferation
Cell Survival
Hydrogel, Polyethylene Glycol Dimethacrylate
Kinetics
Mice
Neurons
Phenotype
Polyethylene Glycols
Reproducibility of Results
Stem Cells
Tissue Array Analysis
English The neurosphere assay is the standard retrospective assay to test the self-renewal capability and multipotency of neural stem cells (NSCs) in vitro. However, it has recently become clear that not all neurospheres are derived from a NSC and that on conventional cell culture substrates, neurosphere motility may cause frequent neurosphere "merging" [Nat Methods 2006;3:801-806; Stem Cells 2007;25:871-874]. Combining biomimetic hydrogel matrix technology with microengineering, we developed a microwell array platform on which NSC fate and neurosphere formation can be unequivocally attributed to a single founding cell. Using time-lapse microscopy and retrospective immunostaining, the fate of several hundred single NSCs was quantified. Compared with conventional neurosphere culture methods on plastic dishes, we detected a more than 100% increase in single NSC viability on soft hydrogels. Effective confinement of single proliferating cells to microwells led to neurosphere formation of vastly different sizes, a high percentage of which showed stem cell phenotypes after one week in culture. The reliability and increased throughput of this platform should help to better elucidate the function of sphere-forming stem/progenitor cells independent of their proliferation dynamics. Disclosure of potential conflicts of interest is found at the end of this article.
Language
  • English
Open access status
green
Identifiers
Persistent URL
https://sonar.ch/global/documents/190212
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