Cellular microarrays for assessing single-cell phenotypic changes in vascular cell populations

Microengineering technologies provide bespoke tools for single-cell studies, including microarray approaches. There are many challenges when culturing adherent single cells in confined geometries for extended periods, including the ability of migratory cells to overcome confining cell-repellent surfaces with time. Following studies suggesting clonal expansion of only a few vascular smooth muscle cells (vSMCs) contributes to plaque formation, the investigation of vSMCs at the single-cell level is central to furthering our understanding of atherosclerosis. Herein, we present a medium throughput cellular microarray, for the tracking of single, freshly-isolated vSMCs as they undergo phenotypic modulation in vitro. Our solution facilitates long-term cell confinement (> 3 weeks) utilising novel application of surface functionalisation methods to define individual culture microwells. We demonstrate successful tracking of hundreds of native vSMCs isolated from rat aortic and carotid artery tissue, monitoring their proliferative capacity and uptake of oxidised low-density lipoprotein (oxLDL) by live-cell microscopy. After 7 days in vitro, the majority of viable SMCs remained as single non-proliferating cells (51% aorta, 78% carotid). However, a sub-population of vSMCs demonstrated high proliferative capacity (≥ 10 progeny; 18% aorta, 5% carotid), in line with reports that a limited number of medial SMCs selectively expand to populate atherosclerotic lesions. Furthermore, we show that, when exposed to oxLDL, proliferative cells uptake higher levels of lipoproteins, whilst also expressing greater levels of galectin-3. Our microwell array approach enables long-term characterisation of multiple phenotypic characteristics and the identification of new cellular sub-populations in migratory, proliferative adherent cell types.

• Publication year

  2023

• Venue

  Biomedical microdevices

• Publication date

  2023-03-16

• Fields of study

  Medicine, Engineering

• Identifiers
  DOI 10.1007/s10544-023-00651-5PMID 36928445PMCID 10020314
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• AI-aided high-throughput profiling of single-cell migration and proliferation on addressable dual-nested microwell arrays

  2023cited by this paper
• Large-scale investigation of single cell activities and response dynamics in a microarray chip with a microfluidics-fabricated microporous membrane.

  2021cited by this paper
• Smooth muscle cells in atherosclerosis: clones but not carbon copies

  2021cited by this paper
• Targeting smooth muscle cell phenotypic switching in vascular disease

  2021cited by this paper
• Vascular smooth muscle cell phenotype switching in carotid atherosclerosis

  2021cited by this paper
• Stem Cell Pluripotency Genes Klf4 and Oct4 Regulate Complex SMC Phenotypic Changes Critical in Late-Stage Atherosclerotic Lesion Pathogenesis

  2020cited by this paper
• Integrated Microfluidic Device for Functional Secretory Immunophenotyping of Immune Cells.

  2020cited by this paper
• Using the Patterned Microarray Culture to Obtain Gene-Editing Monoclonal Cells.

  2020cited by this paper
• Massively parallel quantification of phenotypic heterogeneity in single-cell drug responses

  2020cited by this paper
• 2-Methacryloyloxyethyl Phosphorylcholine Polymer Coating Inhibits Bacterial Adhesion and Biofilm Formation on a Suture: An In Vitro and In Vivo Study

  2020cited by this paper
• Single-cell nucleic acid profiling in droplets (SNAPD) enables high-throughput analysis of heterogeneous cell populations

  2020cited by this paper
• Intestinal Organoid Culture in Polymer Film‐Based Microwell Arrays

  2020cited by this paper
• Automated platform for cell selection and separation based on four-dimensional motility and matrix degradation.

  2019cited by this paper
• Smooth Muscle Cells Contribute the Majority of Foam Cells in ApoE (Apolipoprotein E)-Deficient Mouse Atherosclerosis

  2019cited by this paper
• Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis

  2019cited by this paper
• Critical Areas of Proliferation of Single Cells on Micropatterned Surfaces and Corresponding Cell Type Dependence.

  2019cited by this paper
• Amelioration of the apoptosis-mediated death in isolated human pancreatic islets by minocycline.

  2019cited by this paper
• Drug screening of biopsy-derived spheroids using a self-generated microfluidic concentration gradient

  2018cited by this paper
• Disease-relevant transcriptional signatures identified in individual smooth muscle cells from healthy mouse vessels

  2018cited by this paper
• Smooth muscle cell fate and plasticity in atherosclerosis

  2018cited by this paper
• Single-step fabrication of polydimethylsiloxane microwell arrays with long-lasting hydrophilic inner surfaces

  2018cited by this paper
• Diverse cellular architecture of atherosclerotic plaque derives from clonal expansion of a few medial SMCs.

  2017cited by this paper
• Extensive Proliferation of a Subset of Differentiated, yet Plastic, Medial Vascular Smooth Muscle Cells Contributes to Neointimal Formation in Mouse Injury and Atherosclerosis Models

  2016cited by this paper
• Vascular Smooth Muscle Cells in Atherosclerosis.

  2016cited by this paper
• The transition of smooth muscle cells from a contractile to a migratory, phagocytic phenotype: direct demonstration of phenotypic modulation

  2016cited by this paper
• Differential regulation of caspase-2 in MPP⁺-induced apoptosis in primary cortical neurons.

  2015cited by this paper
• KLF4 Dependent Phenotypic Modulation of SMCs Plays a Key Role in Atherosclerotic Plaque Pathogenesis

  2015cited by this paper
• Programmable Laser-Assisted Surface Microfabrication on a Poly(Vinyl Alcohol)-Coated Glass Chip with Self-Changing Cell Adhesivity for Heterotypic Cell Patterning.

  2015cited by this paper
• Microwell arrays reveal cellular heterogeneity during the clonal expansion of transformed human cells.

  2015cited by this paper
• Ligustrazine attenuates the platelet-derived growth factor-BB-induced proliferation and migration of vascular smooth muscle cells by interrupting extracellular signal-regulated kinase and P38 mitogen-activated protein kinase pathways.

  2015cited by this paper
• Fabrication of a cell-adhesive microwell array for 3-dimensional in vitro cell model

  2015cited by this paper
• Collagen surfaces modified with photo-cleavable polyethylene glycol-lipid support versatile single-cell arrays of both non-adherent and adherent cells.

  2014cited by this paper
• Primary hepatocytes and their cultures in liver apoptosis research

  2014cited by this paper
• Transdifferentiation of Vascular Smooth Muscle Cells to Macrophage-Like Cells During Atherogenesis

  2014cited by this paper
• Previously differentiated medial vascular smooth muscle cells contribute to neointima formation following vascular injury

  2014cited by this paper
• Monitoring the dynamics of primary T cell activation and differentiation using long term live cell imaging in microwell arrays.

  2012cited by this paper
• High-throughput clonal analysis of neural stem cells in microarrayed artificial niches.

  2012cited by this paper
• Detection of Histone Modifications at Specific Gene Loci in Single Cells in Histological Sections

  2012cited by this paper
• A silicon-glass microwell platform for high-resolution imaging and high-content screening with single cell resolution

  2011cited by this paper
• Dynamics of galectin-3 in the nucleus and cytoplasm.

  2010cited by this paper
• Enhancing the Reliability and Throughput of Neurosphere Culture on Hydrogel Microwell Arrays

  2008cited by this paper
• Micropallet arrays with poly(ethylene glycol) walls.

  2008cited by this paper
• single cells{

  2007cited by this paper
• High-resolution video monitoring of hematopoietic stem cells cultured in single-cell arrays identifies new features of self-renewal.

  2006cited by this paper
• Large-scale single-cell trapping and imaging using microwell arrays.

  2005cited by this paper
• Simple Approach to Micropattern Cells on Common Culture Substrates by Tuning Substrate Wettability

  2004cited by this paper
• "Microcanals" for micropipette access to single cells in microfluidic environments.

  2004cited by this paper
• Transdifferentiation of mouse aortic smooth muscle cells to a macrophage-like state after cholesterol loading

  2003cited by this paper
• Nuclear localisation of wild type and mutant galectin‐3 in transfected cells

  2000cited by this paper

• No citing papers are available for this paper.