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Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting
BACKGROUND: Molecular characterisation of single circulating tumour cells (CTCs) holds considerable promise for predictive biomarker assessment and to explore CTC heterogeneity. We evaluate a new method, the DEPArray system, that allows the dielectrophoretic manipulation and isolation of single and...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3619252/ https://www.ncbi.nlm.nih.gov/pubmed/23470469 http://dx.doi.org/10.1038/bjc.2013.92 |
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author | Peeters, D J E De Laere, B Van den Eynden, G G Van Laere, S J Rothé, F Ignatiadis, M Sieuwerts, A M Lambrechts, D Rutten, A van Dam, P A Pauwels, P Peeters, M Vermeulen, P B Dirix, L Y |
author_facet | Peeters, D J E De Laere, B Van den Eynden, G G Van Laere, S J Rothé, F Ignatiadis, M Sieuwerts, A M Lambrechts, D Rutten, A van Dam, P A Pauwels, P Peeters, M Vermeulen, P B Dirix, L Y |
author_sort | Peeters, D J E |
collection | PubMed |
description | BACKGROUND: Molecular characterisation of single circulating tumour cells (CTCs) holds considerable promise for predictive biomarker assessment and to explore CTC heterogeneity. We evaluate a new method, the DEPArray system, that allows the dielectrophoretic manipulation and isolation of single and 100% purified groups of CTCs from pre-enriched blood samples and explore the feasibility of their molecular characterisation. METHODS: Samples containing known numbers of two cell populations were used to assess cell loss during sample loading. Cultured breast cancer cells were isolated from spiked blood samples using CellSearch CTC and Profile kits. Single tumour cells and groups of up to 10 tumour cells were recovered with the DEPArray system and subjected to transcriptional and mutation analysis. RESULTS: On average, 40% cell loss was observed when loading samples to the DEPArray system. Expected mutations in clinically relevant markers could be obtained for 60% of single recovered tumour cells and all groups of tumour cells. Reliable gene expression profiles were obtained from single cells and groups of up to 10 cells for 2 out of 3 spiked breast cancer cell lines. CONCLUSION: We describe a semiautomated workflow for the isolation of small groups of 1 to 10 tumour cells from whole blood samples and provide proof of principle for the feasibility of their comprehensive molecular characterisation. |
format | Online Article Text |
id | pubmed-3619252 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-36192522014-04-02 Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting Peeters, D J E De Laere, B Van den Eynden, G G Van Laere, S J Rothé, F Ignatiadis, M Sieuwerts, A M Lambrechts, D Rutten, A van Dam, P A Pauwels, P Peeters, M Vermeulen, P B Dirix, L Y Br J Cancer Molecular Diagnostics BACKGROUND: Molecular characterisation of single circulating tumour cells (CTCs) holds considerable promise for predictive biomarker assessment and to explore CTC heterogeneity. We evaluate a new method, the DEPArray system, that allows the dielectrophoretic manipulation and isolation of single and 100% purified groups of CTCs from pre-enriched blood samples and explore the feasibility of their molecular characterisation. METHODS: Samples containing known numbers of two cell populations were used to assess cell loss during sample loading. Cultured breast cancer cells were isolated from spiked blood samples using CellSearch CTC and Profile kits. Single tumour cells and groups of up to 10 tumour cells were recovered with the DEPArray system and subjected to transcriptional and mutation analysis. RESULTS: On average, 40% cell loss was observed when loading samples to the DEPArray system. Expected mutations in clinically relevant markers could be obtained for 60% of single recovered tumour cells and all groups of tumour cells. Reliable gene expression profiles were obtained from single cells and groups of up to 10 cells for 2 out of 3 spiked breast cancer cell lines. CONCLUSION: We describe a semiautomated workflow for the isolation of small groups of 1 to 10 tumour cells from whole blood samples and provide proof of principle for the feasibility of their comprehensive molecular characterisation. Nature Publishing Group 2013-04-02 2013-03-07 /pmc/articles/PMC3619252/ /pubmed/23470469 http://dx.doi.org/10.1038/bjc.2013.92 Text en Copyright © 2013 Cancer Research UK http://creativecommons.org/licenses/by-nc-sa/3.0/ From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/ |
spellingShingle | Molecular Diagnostics Peeters, D J E De Laere, B Van den Eynden, G G Van Laere, S J Rothé, F Ignatiadis, M Sieuwerts, A M Lambrechts, D Rutten, A van Dam, P A Pauwels, P Peeters, M Vermeulen, P B Dirix, L Y Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting |
title | Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting |
title_full | Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting |
title_fullStr | Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting |
title_full_unstemmed | Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting |
title_short | Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting |
title_sort | semiautomated isolation and molecular characterisation of single or highly purified tumour cells from cellsearch enriched blood samples using dielectrophoretic cell sorting |
topic | Molecular Diagnostics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3619252/ https://www.ncbi.nlm.nih.gov/pubmed/23470469 http://dx.doi.org/10.1038/bjc.2013.92 |
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