A novel method for the isolation of single cells mimicking circulating tumour cells adhered on Smart Bio Surface slides by Laser Capture Microdissection

In recent years, the importance of isolating single cells from blood circulation for several applications, such as non-invasive tumour diagnosis, the monitoring of minimal residual disease, and the analysis of circulating fetal cells for prenatal diagnosis, urged the need to set up innovative method...

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Veröffentlicht in:PloS one 2024-03, Vol.19 (3), p.e0297739-e0297739
Hauptverfasser: Visci, Grazia, Tolomeo, Doron, Lonoce, Angelo, Arshadi, Aram, Bascetta, Lorenzo, Trotta, Gianluca, van Riel, Margot, Vermeesch, Joris Robert, Carbone, Roberta, Storlazzi, Clelia Tiziana
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container_title PloS one
container_volume 19
creator Visci, Grazia
Tolomeo, Doron
Lonoce, Angelo
Arshadi, Aram
Bascetta, Lorenzo
Trotta, Gianluca
van Riel, Margot
Vermeesch, Joris Robert
Carbone, Roberta
Storlazzi, Clelia Tiziana
description In recent years, the importance of isolating single cells from blood circulation for several applications, such as non-invasive tumour diagnosis, the monitoring of minimal residual disease, and the analysis of circulating fetal cells for prenatal diagnosis, urged the need to set up innovative methods. For such applications, different methods were developed. All show some weaknesses, especially a limited sensitivity, and specificity. Here we present a new method for isolating a single or a limited number of cells adhered to SBS slides (Tethis S.p.a.) (a glass slide coated with Nanostructured Titanium Dioxide) by Laser Capture Microdissection (LCM) and subsequent Whole Genome Amplification. SBS slides have been shown to have an optimal performance in immobilizing circulating tumour cells (CTCs) from early breast cancer patients. In this work, we spiked cancer cells in blood samples to mimic CTCs. By defining laser parameters to cut intact samples, we were able to isolate genetically intact single cells. We demonstrate that SBS slides are optimally suited for isolating cells using LCM and that this method provides high-quality DNA, ideal for gene-specific assays such as PCR and Sanger sequencing for mutation analysis.
doi_str_mv 10.1371/journal.pone.0297739
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subjects Analysis
B cells
Cancer
Cancer cells
Care and treatment
Diagnosis
DNA sequencing
Ethylenediaminetetraacetic acid
Gene mutations
Genomes
Genomics
Health aspects
Metastasis
Methods
Nucleotide sequencing
Tumors
title A novel method for the isolation of single cells mimicking circulating tumour cells adhered on Smart Bio Surface slides by Laser Capture Microdissection
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