High-throughput combinatorial screening reveals interactions between signaling molecules that regulate adult neural stem cell fate

Riya Muckom, Sean McFarland, Chun Yang, Brian Perea, Megan Gentes, Abirami Murugappan, Eric Tran, Jonathan S. Dordick, Douglas S. Clark, David V. Schaffer

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Advancing our knowledge of how neural stem cell (NSC) behavior in the adult hippocampus is regulated has implications for elucidating basic mechanisms of learning and memory as well as for neurodegenerative disease therapy. To date, numerous biochemical cues from the endogenous hippocampal NSC niche have been identified as modulators of NSC quiescence, proliferation, and differentiation; however, the complex repertoire of signaling factors within stem cell niches raises the question of how cues act in combination with one another to influence NSC physiology. To help overcome experimental bottlenecks in studying this question, we adapted a high-throughput microculture system, with over 500 distinct microenvironments, to conduct a systematic combinatorial screen of key signaling cues and collect high-content phenotype data on endpoint NSC populations. This novel application of the platform consumed only 0.2% of reagent volumes used in conventional 96-well plates, and resulted in the discovery of numerous statistically significant interactions among key endogenous signals. Antagonistic relationships between fibroblast growth factor 2, transforming growth factor β (TGF-β), and Wnt-3a were found to impact NSC proliferation and differentiation, whereas a synergistic relationship between Wnt-3a and Ephrin-B2 on neuronal differentiation and maturation was found. Furthermore, TGF-β and bone morphogenetic protein 4 combined with Wnt-3a and Ephrin-B2 resulted in a coordinated effect on neuronal differentiation and maturation. Overall, this study offers candidates for further elucidation of significant mechanisms guiding NSC fate choice and contributes strategies for enhancing control over stem cell-based therapies for neurodegenerative diseases.

Original languageEnglish
Pages (from-to)193-205
Number of pages13
JournalBiotechnology and Bioengineering
Volume116
Issue number1
DOIs
StatePublished - Jan 2019
Externally publishedYes

Keywords

  • NSC
  • combinatorial
  • high-throughput
  • hippocampus
  • niche

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