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Dynamic gene expression profiling using a microfabricated living cell array.

Deanna M Thompson1, Kevin R King, Kenneth J Wieder

  • 1Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospitals for Children, Boston, Massachusetts 02114, USA.

Analytical Chemistry
|July 16, 2004
PubMed
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We developed a living cell array (LCA) microfluidic platform for real-time gene expression monitoring. This system enables high-throughput analysis of cellular responses to molecular stimuli, advancing our understanding of dynamic biological processes.

Area of Science:

  • Cell Biology
  • Biotechnology
  • Microfluidics

Background:

  • Accurate monitoring of gene expression dynamics in live cells is crucial for understanding cellular responses.
  • Existing methods often lack the throughput or non-destructive capabilities for comprehensive dynamic analysis.

Purpose of the Study:

  • To develop a microfluidic platform for continuous, high-throughput, and nondestructive monitoring of gene expression in individual live cells.
  • To create a 'living cell array' (LCA) capable of integrating molecular stimulation with precise measurement of cellular responses.

Main Methods:

  • Development of an optically transparent microfluidic device integrating upstream molecular stimulus generation with downstream reporter cell stimulation.
  • Utilizing reporter cells with green fluorescence plasmids to quantify gene expression dynamics.

Related Experiment Videos

  • Continuous monitoring and quantification of cellular fluorescence to infer gene expression patterns.

    • Main Results:

    • Demonstrated the LCA platform's capability by profiling the activation of the transcription factor NF-kappaB in HeLa S3 cells.
    • Successfully correlated varying doses of TNF-alpha with specific gene expression dynamics.
    • Showcased the platform's ability to control dynamic inputs and measure dynamic outputs from adherent mammalian cells.

    Conclusions:

    • The living cell array (LCA) platform provides a novel method for high-throughput profiling of gene expression dynamics.
    • This approach facilitates a deeper understanding of cellular responses to soluble stimuli.
    • Integration with techniques like DNA microarrays offers a comprehensive view of dynamic cellular signaling pathways.