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A digital microfluidic platform for primary cell culture and analysis.

Suthan Srigunapalan1, Irwin A Eydelnant, Craig A Simmons

  • 1Department of Mechanical & Industrial Engineering, University of Toronto, 5 King's College Road, Toronto, Ontario, Canada M5S 3G8.

Lab on a Chip
|November 19, 2011
PubMed
Summary

Digital microfluidics (DMF) enables primary cell culture and analysis. This new method successfully cultured diverse primary cells and demonstrated their in vivo-like functionality for research applications.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Cell Biology

Background:

  • Digital microfluidics (DMF) offers advantages like automation and parallelization for sample analysis.
  • Previous DMF applications primarily used immortalized cell lines, limiting its use with primary cells.
  • There is a growing need for microfluidic platforms capable of handling primary cell cultures.

Purpose of the Study:

  • To develop and validate the first digital microfluidic (DMF) method for primary cell culture and analysis.
  • To demonstrate the feasibility of an "upside-down" cell culture technique in DMF devices.
  • To evaluate the functionality and in vivo-like properties of primary cells cultured using DMF.

Main Methods:

  • Developed a novel "upside-down" cell culture method by patterning the top plate of a DMF device.

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  • Cultured three distinct primary cell types for up to one week within the DMF platform.
  • Implemented fixation, permeabilization, and staining protocols for F-actin and nuclei.
  • Performed a multistep assay to assess monocyte adhesion to endothelial cells (ECs) in DMF.

    • Main Results:

    • Successfully cultured three primary cell types for seven days using the "upside-down" DMF method.
    • Demonstrated successful co-culture and functional assays, including monocyte-endothelial cell adhesion.
    • Observed significantly higher monocyte adhesion to TNF-α-stimulated ECs, indicating preserved in vivo-like functionality.
    • Validated the utility of DMF for manipulating, maintaining, and analyzing primary cells.

    Conclusions:

    • The developed "upside-down" digital microfluidic method is effective for culturing and analyzing primary cells.
    • DMF platforms can maintain the in vivo-like properties and functionality of primary cells.
    • This technology offers a valuable tool for research involving precious primary cell samples from small animals or patients.