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Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology
There has been increased interest in utilizing non-invasive “liquid biopsies” to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both ge...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5859469/ https://www.ncbi.nlm.nih.gov/pubmed/29872702 http://dx.doi.org/10.1038/s41698-017-0015-0 |
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author | Renier, Corinne Pao, Edward Che, James Liu, Haiyan E. Lemaire, Clementine A. Matsumoto, Melissa Triboulet, Melanie Srivinas, Sandy Jeffrey, Stefanie S. Rettig, Matthew Kulkarni, Rajan P. Di Carlo, Dino Sollier-Christen, Elodie |
author_facet | Renier, Corinne Pao, Edward Che, James Liu, Haiyan E. Lemaire, Clementine A. Matsumoto, Melissa Triboulet, Melanie Srivinas, Sandy Jeffrey, Stefanie S. Rettig, Matthew Kulkarni, Rajan P. Di Carlo, Dino Sollier-Christen, Elodie |
author_sort | Renier, Corinne |
collection | PubMed |
description | There has been increased interest in utilizing non-invasive “liquid biopsies” to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on high capture efficiency but with lower purity and often long and manual sample preparation, which limits the use of captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can capture CTCs with high purity (from 1.74 to 37.59%) and efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46–77 years old; 1.25–2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21–27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial–mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples. |
format | Online Article Text |
id | pubmed-5859469 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-58594692018-06-05 Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology Renier, Corinne Pao, Edward Che, James Liu, Haiyan E. Lemaire, Clementine A. Matsumoto, Melissa Triboulet, Melanie Srivinas, Sandy Jeffrey, Stefanie S. Rettig, Matthew Kulkarni, Rajan P. Di Carlo, Dino Sollier-Christen, Elodie NPJ Precis Oncol Article There has been increased interest in utilizing non-invasive “liquid biopsies” to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on high capture efficiency but with lower purity and often long and manual sample preparation, which limits the use of captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can capture CTCs with high purity (from 1.74 to 37.59%) and efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46–77 years old; 1.25–2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21–27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial–mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples. Nature Publishing Group UK 2017-05-08 /pmc/articles/PMC5859469/ /pubmed/29872702 http://dx.doi.org/10.1038/s41698-017-0015-0 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Renier, Corinne Pao, Edward Che, James Liu, Haiyan E. Lemaire, Clementine A. Matsumoto, Melissa Triboulet, Melanie Srivinas, Sandy Jeffrey, Stefanie S. Rettig, Matthew Kulkarni, Rajan P. Di Carlo, Dino Sollier-Christen, Elodie Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology |
title | Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology |
title_full | Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology |
title_fullStr | Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology |
title_full_unstemmed | Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology |
title_short | Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology |
title_sort | label-free isolation of prostate circulating tumor cells using vortex microfluidic technology |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5859469/ https://www.ncbi.nlm.nih.gov/pubmed/29872702 http://dx.doi.org/10.1038/s41698-017-0015-0 |
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