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A High-throughput Cell Microarray Platform for Correlative Analysis of Cell Differentiation and Traction Forces
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Cellular microarrays for assessing single-cell phenotypic changes in vascular cell populations.

E Smith1,2, M Zagnoni1, M E Sandison3

  • 1Electronic & Electrical Engineering, Royal College Building, University of Strathclyde, G1 1XW, Glasgow, UK.

Biomedical Microdevices
|March 17, 2023
PubMed
Summary

A novel cellular microarray enables long-term tracking of single vascular smooth muscle cells (vSMCs), revealing subpopulations with high proliferative capacity that contribute to atherosclerosis plaque formation and oxLDL uptake.

Keywords:
Microwell arrayPhenotypic modulationSingle cellSmooth muscleoxLDL

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Cardiovascular Research

Background:

  • Single-cell studies are crucial for understanding complex biological processes like atherosclerosis.
  • Culturing adherent single cells long-term presents challenges, particularly with migratory cells and specialized surfaces.
  • Vascular smooth muscle cells (vSMCs) clonal expansion is implicated in atherosclerotic plaque development.

Purpose of the Study:

  • To develop and validate a medium-throughput cellular microarray for tracking individual, freshly-isolated vSMCs in vitro.
  • To investigate vSMC phenotypic modulation, proliferative capacity, and oxidized low-density lipoprotein (oxLDL) uptake over extended culture periods.
  • To identify distinct vSMC subpopulations involved in atherosclerosis pathogenesis.

Main Methods:

  • Utilized microengineering to create a cellular microarray with novel surface functionalization for long-term single-cell confinement (>3 weeks).
  • Isolated and cultured native vSMCs from rat aortic and carotid artery tissues within individual microwells.
  • Employed live-cell microscopy to monitor vSMC proliferation and oxLDL uptake, alongside galectin-3 expression.

Main Results:

  • Successfully confined and tracked hundreds of single vSMCs, demonstrating the microarray's efficacy.
  • Identified a sub-population of vSMCs exhibiting high proliferative capacity (≥10 progeny), consistent with roles in atherosclerotic lesion expansion.
  • Showed that proliferative vSMCs have increased oxLDL uptake and higher galectin-3 expression when exposed to oxLDL.

Conclusions:

  • The developed microwell array facilitates long-term characterization of single adherent cells, enabling the identification of functionally distinct subpopulations.
  • This approach provides new insights into vSMC heterogeneity and their specific contributions to atherosclerosis.
  • The technology supports the study of migratory and proliferative cell types, advancing our understanding of cardiovascular disease mechanisms.