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A Microfluidic Platform for High-throughput Single-cell Isolation and Culture
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Microfluidic platform accelerates tissue processing into single cells for molecular analysis and primary culture

Jeremy A Lombardo1, Marzieh Aliaghaei2, Quy H Nguyen3

  • 1Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, USA.

Nature Communications
|May 18, 2021
PubMed
Summary

This study introduces a microfluidic platform for efficient tissue dissociation, yielding more single cells and reducing processing time. The technology enhances cell recovery and reproducibility for single-cell analysis.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microfluidics

Background:

  • Single-cell analysis requires efficient tissue dissociation into single-cell suspensions.
  • Current dissociation methods are often inefficient, labor-intensive, variable, and biased.
  • This limits the characterization of complex tissue heterogeneity.

Purpose of the Study:

  • To develop and evaluate a novel microfluidic platform for tissue processing.
  • To improve the efficiency, reproducibility, and cell yield of single-cell suspensions.
  • To enable unbiased characterization of diverse cell subtypes within tissues.

Main Methods:

  • A microfluidic platform integrating tissue digestion, disaggregation, and filtration was developed.
  • The platform was tested on kidney, mammary tumor, liver, and heart tissues.
  • Single cell RNA sequencing was used to assess cell populations and stress responses.

Main Results:

  • Microfluidic processing increased single-cell yield by 2.5-fold for kidney and mammary tumor.
  • Endothelial cells, fibroblasts, and basal epithelium were enriched without increased stress.
  • Processing time was significantly reduced for liver and heart tissues.
  • Periodic cell recovery increased hepatocyte and cardiomyocyte numbers and batch-to-batch reproducibility.

Conclusions:

  • The microfluidic platform offers a more efficient and reproducible method for single-cell suspension preparation.
  • This technology facilitates deeper insights into tissue complexity and cellular heterogeneity.
  • The platform has broad applicability across various tissue types for single-cell omics studies.