A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer

Circulating tumor cells (CTCs) have potential as diagnostic, prognostic, and predictive biomarkers in solid tumors. Despite Food and Drug Administration (FDA) approval of CTC devices in various cancers, the rarity and heterogeneity of CTCs in lung cancer make them technically challenging to isolate...

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Main Authors: Volga M Saini, Ezgi Oner, Mark P. Ward, Sinead Hurley, Brian David Henderson, Faye Lewis, Stephen P. Finn, Gerard J. Fitzmaurice, John J. O'Leary, Sharon O'Toole, Lorraine O'Driscoll, Kathy Gately
Format: Article
Language:English
Published: Wiley 2025-07-01
Series:Molecular Oncology
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Online Access:https://doi.org/10.1002/1878-0261.13705
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author Volga M Saini
Ezgi Oner
Mark P. Ward
Sinead Hurley
Brian David Henderson
Faye Lewis
Stephen P. Finn
Gerard J. Fitzmaurice
John J. O'Leary
Sharon O'Toole
Lorraine O'Driscoll
Kathy Gately
author_facet Volga M Saini
Ezgi Oner
Mark P. Ward
Sinead Hurley
Brian David Henderson
Faye Lewis
Stephen P. Finn
Gerard J. Fitzmaurice
John J. O'Leary
Sharon O'Toole
Lorraine O'Driscoll
Kathy Gately
author_sort Volga M Saini
collection DOAJ
description Circulating tumor cells (CTCs) have potential as diagnostic, prognostic, and predictive biomarkers in solid tumors. Despite Food and Drug Administration (FDA) approval of CTC devices in various cancers, the rarity and heterogeneity of CTCs in lung cancer make them technically challenging to isolate and analyze, hindering their clinical integration. Establishing a consensus through comparative analysis of different CTC systems is warranted. This study aimed to evaluate seven different CTC enrichment methods across five technologies using a standardized spike‐in protocol: the CellMag™ (EpCAM‐dependent enrichment), EasySep™ and RosetteSep™ (blood cell depletion), and the Parsortix® PR1 and the new design Parsortix® Prototype (PP) (size‐ and deformability‐based enrichment). The Parsortix® systems were also evaluated for any differences in recovery rates between cell harvest versus in‐cassette staining. Healthy donor blood (5 mL) was spiked with 100 fluorescently labeled EpCAMhigh H1975 cells, processed through each system, and the isolation efficiency was calculated. The CellMag™ had the highest recovery rate (70 ± 14%), followed by Parsortix® PR1 in‐cassette staining, while the EasySep™ had the lowest recovery (18 ± 8%). Additional spike‐in experiments were performed with EpCAMmoderate A549 and EpCAMlow H1299 cells using the CellMag™ and Parsortix® PR1 in‐cassette staining. The recovery rate of CellMag™ significantly reduced to 35 ± 14% with A549 cells and 1 ± 1% with H1299 cells. However, the Parsortix® PR1 in‐cassette staining showed cell phenotype‐independent and consistent recovery rates among all lung cancer cell lines: H1975 (49 ± 2%), A549 (47 ± 10%), and H1299 (52 ± 10%). Furthermore, we demonstrated that the Parsortix® PR1 in‐cassette staining method is capable of isolating heterogeneous single CTCs and cell clusters from patient samples. The Parsortix® PR1 in‐cassette staining, capable of isolating different phenotypes of CTCs as either single cells or cell clusters with consistent recovery rates, is considered optimal for CTC enrichment for lung cancer, albeit needing further optimization and validation.
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spelling doaj-art-0b788ee647874946a0ef4b2db05d92902025-08-20T03:17:43ZengWileyMolecular Oncology1574-78911878-02612025-07-011972014203710.1002/1878-0261.13705A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancerVolga M Saini0Ezgi Oner1Mark P. Ward2Sinead Hurley3Brian David Henderson4Faye Lewis5Stephen P. Finn6Gerard J. Fitzmaurice7John J. O'Leary8Sharon O'Toole9Lorraine O'Driscoll10Kathy Gately11Thoracic Oncology Research Group, Trinity Translational Medicine Institute St James's Hospital Dublin IrelandThoracic Oncology Research Group, Trinity Translational Medicine Institute St James's Hospital Dublin IrelandTrinity St. James's Cancer Institute Trinity College Dublin IrelandThoracic Oncology Research Group, Trinity Translational Medicine Institute St James's Hospital Dublin IrelandTrinity St. James's Cancer Institute Trinity College Dublin IrelandTrinity St. James's Cancer Institute Trinity College Dublin IrelandThoracic Oncology Research Group, Trinity Translational Medicine Institute St James's Hospital Dublin IrelandDepartment of Cardiothoracic Surgery St James's Hospital Dublin IrelandTrinity St. James's Cancer Institute Trinity College Dublin IrelandTrinity St. James's Cancer Institute Trinity College Dublin IrelandTrinity St. James's Cancer Institute Trinity College Dublin IrelandThoracic Oncology Research Group, Trinity Translational Medicine Institute St James's Hospital Dublin IrelandCirculating tumor cells (CTCs) have potential as diagnostic, prognostic, and predictive biomarkers in solid tumors. Despite Food and Drug Administration (FDA) approval of CTC devices in various cancers, the rarity and heterogeneity of CTCs in lung cancer make them technically challenging to isolate and analyze, hindering their clinical integration. Establishing a consensus through comparative analysis of different CTC systems is warranted. This study aimed to evaluate seven different CTC enrichment methods across five technologies using a standardized spike‐in protocol: the CellMag™ (EpCAM‐dependent enrichment), EasySep™ and RosetteSep™ (blood cell depletion), and the Parsortix® PR1 and the new design Parsortix® Prototype (PP) (size‐ and deformability‐based enrichment). The Parsortix® systems were also evaluated for any differences in recovery rates between cell harvest versus in‐cassette staining. Healthy donor blood (5 mL) was spiked with 100 fluorescently labeled EpCAMhigh H1975 cells, processed through each system, and the isolation efficiency was calculated. The CellMag™ had the highest recovery rate (70 ± 14%), followed by Parsortix® PR1 in‐cassette staining, while the EasySep™ had the lowest recovery (18 ± 8%). Additional spike‐in experiments were performed with EpCAMmoderate A549 and EpCAMlow H1299 cells using the CellMag™ and Parsortix® PR1 in‐cassette staining. The recovery rate of CellMag™ significantly reduced to 35 ± 14% with A549 cells and 1 ± 1% with H1299 cells. However, the Parsortix® PR1 in‐cassette staining showed cell phenotype‐independent and consistent recovery rates among all lung cancer cell lines: H1975 (49 ± 2%), A549 (47 ± 10%), and H1299 (52 ± 10%). Furthermore, we demonstrated that the Parsortix® PR1 in‐cassette staining method is capable of isolating heterogeneous single CTCs and cell clusters from patient samples. The Parsortix® PR1 in‐cassette staining, capable of isolating different phenotypes of CTCs as either single cells or cell clusters with consistent recovery rates, is considered optimal for CTC enrichment for lung cancer, albeit needing further optimization and validation.https://doi.org/10.1002/1878-0261.13705CellMag™circulating tumor cellsEasySep™non–small cell lung cancerParsortix®RosetteSep™
spellingShingle Volga M Saini
Ezgi Oner
Mark P. Ward
Sinead Hurley
Brian David Henderson
Faye Lewis
Stephen P. Finn
Gerard J. Fitzmaurice
John J. O'Leary
Sharon O'Toole
Lorraine O'Driscoll
Kathy Gately
A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
Molecular Oncology
CellMag™
circulating tumor cells
EasySep™
non–small cell lung cancer
Parsortix®
RosetteSep™
title A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
title_full A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
title_fullStr A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
title_full_unstemmed A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
title_short A comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
title_sort comparative study of circulating tumor cell isolation and enumeration technologies in lung cancer
topic CellMag™
circulating tumor cells
EasySep™
non–small cell lung cancer
Parsortix®
RosetteSep™
url https://doi.org/10.1002/1878-0261.13705
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