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A Microfluidics Approach for the Functional Investigation of Signaling Oscillations Governing Somitogenesis
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Microfluidic perfusion for regulating diffusible signaling in stem cells.

Katarina Blagovic1, Lily Y Kim, Joel Voldman

  • 1Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.

Plos One
|August 11, 2011
PubMed
Summary
This summary is machine-generated.

Microfluidic platforms can remove cell-secreted factors to study autocrine signaling in embryonic stem cells (ESCs). This reveals FGF4-dependent and -independent pathways crucial for neuroectodermal commitment.

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Area of Science:

  • Cell Biology
  • Developmental Biology
  • Microfluidics

Background:

  • Autocrine and paracrine signaling regulate embryonic stem cell (ESC) pluripotency and differentiation.
  • Fibroblast growth factor-4 (FGF4) autocrine signaling is vital for mouse ESC (mESC) neuroectodermal specification.
  • Controlling diffusible signaling in vitro is challenging due to its confined nature.

Purpose of the Study:

  • To investigate the role of autocrine signaling in mESC neuroectodermal specification.
  • To develop a novel microfluidic approach to experimentally control and downregulate diffusible signaling.
  • To determine if FGF4 signaling alone is sufficient for neuroectodermal fate or if other factors are involved.

Main Methods:

  • Development of a multiplex microfluidic platform for continuous removal of cell-secreted factors.
  • Comparison of cell growth and differentiation in chambers with and without secreted factors to isolate signaling effects.
  • Induction of FGF4 signaling and inhibition of FGF signaling in supplemented media to identify non-FGF factors.

Main Results:

  • Cell-secreted growth factors are essential for mESC neuroectodermal specification.
  • Non-FGF cell-secreted factors are required to support the growth of differentiating mESCs.
  • Microfluidic perfusion effectively downregulates autocrine/paracrine signaling, enabling study of diffusible factor sufficiency.

Conclusions:

  • Microfluidic perfusion can downregulate autocrine/paracrine signaling, allowing examination of extracellular factor sufficiency.
  • Autocrine/paracrine signaling drives mESC neuroectodermal commitment via both FGF4-dependent and -independent pathways.
  • This microfluidic technique uncovers previously hidden autocrine/paracrine processes, establishing it as a tool for studying diffusible signaling.