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Related Experiment Videos

Microfluidic arrays for logarithmically perfused embryonic stem cell culture.

Lily Kim1, Michael D Vahey, Hsu-Yi Lee

  • 1Massachusetts Institute of Technology, 77 Massachusetts Avenue, Rm 36-824, Cambridge, MA 02139, USA.

Lab on a Chip
|March 3, 2006
PubMed
Summary

This study introduces a novel microfluidic device for culturing cells under precisely controlled, logarithmically scaled flow rates. This technology enhances control over the cellular microenvironment, crucial for sensitive cell types like embryonic stem cells.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Cell Biology

Background:

  • Static cell culture limits control over the soluble microenvironment.
  • Embryonic stem cells (ESCs) are highly sensitive to microenvironmental conditions.
  • Perfusion culture offers enhanced control but often lacks precise flow rate scaling.

Purpose of the Study:

  • To develop and characterize a microfluidic device for culturing adherent cells across a logarithmic range of flow rates.
  • To demonstrate the device's capability for creating logarithmic concentration gradients.
  • To investigate the effects of controlled perfusion on murine embryonic stem cells (mESCs) and 3T3 fibroblasts.

Main Methods:

  • Fabrication of a microfluidic device with integrated fluidic resistances for syringe-driven flow.

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  • Characterization of flow rates using particle velocimetry.
  • Extension of the device to generate logarithmic concentration gradients, verified by fluorescence measurements.
  • Culturing of mESCs and 3T3 fibroblasts under continuous, logarithmically scaled perfusion.

    • Main Results:

    • The microfluidic device successfully established logarithmic flow rates across four chambers (>300x range).
    • An extended device design created verified logarithmic concentration gradients.
    • mESCs cultured for 4 days showed healthy morphology at higher flow rates, with no proliferation at the slowest rate.
    • 3T3 fibroblasts cultured for 3 days proliferated at all but the slowest flow rates.

    Conclusions:

    • The developed microfluidic device provides precise control over the cellular microenvironment through scalable, logarithmic perfusion.
    • This technology is particularly beneficial for culturing sensitive cells like ESCs, enabling detailed study of flow-dependent behaviors.
    • The device facilitates enhanced control over molecular transport, advancing cell culture methodologies.