IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells

Background: detailed information about circulating tumor cells (CTCs) as an indicator of therapy response and cancer metastasis is crucial not only for basic research but also for diagnostics and therapeutic approaches. Here, we showcase a newly developed IsoMAG IMS system with an optimized protocol...

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Autores principales:	Alena Gribko, Janis Stiefel, Lana Liebetanz, Sophie Madeleine Nagel, Julian Künzel, Madita Wandrey, Jan Hagemann, Roland H. Stauber, Christian Freese, Désirée Gül
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	automation liquid biopsy circulating tumor cells head and neck squamous cell carcinoma immunomagnetic particle-based detection metastasis Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/530d9fbd81f045eba42d343f7ae680f5
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spelling	oai:doaj.org-article:530d9fbd81f045eba42d343f7ae680f52021-11-25T17:21:04ZIsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells10.3390/diagnostics111120402075-4418https://doaj.org/article/530d9fbd81f045eba42d343f7ae680f52021-11-01T00:00:00Zhttps://www.mdpi.com/2075-4418/11/11/2040https://doaj.org/toc/2075-4418Background: detailed information about circulating tumor cells (CTCs) as an indicator of therapy response and cancer metastasis is crucial not only for basic research but also for diagnostics and therapeutic approaches. Here, we showcase a newly developed IsoMAG IMS system with an optimized protocol for fully automated immunomagnetic enrichment of CTCs, also revealing rare CTC subpopulations. Methods: using different squamous cell carcinoma cell lines, we developed an isolation protocol exploiting highly efficient EpCAM-targeting magnetic beads for automated CTC enrichment by the IsoMAG IMS system. By FACS analysis, we analyzed white blood contamination usually preventing further downstream analysis of enriched cells. Results: 1 µm magnetic beads with tosyl-activated hydrophobic surface properties were found to be optimal for automated CTC enrichment. More than 86.5% and 95% of spiked cancer cells were recovered from both cell culture media or human blood employing our developed protocol. In addition, contamination with white blood cells was minimized to about 1200 cells starting from 7.5 mL blood. Finally, we showed that the system is applicable for HNSCC patient samples and characterized isolated CTCs by immunostaining using a panel of tumor markers. Conclusion: Herein, we demonstrate that the IsoMAG system allows the detection and isolation of CTCs from HNSCC patient blood for disease monitoring in a fully-automated process with a significant leukocyte count reduction. Future developments seek to integrate the IsoMAG IMS system into an automated microfluidic-based isolation workflow to further facilitate single CTC detection also in clinical routine.Alena GribkoJanis StiefelLana LiebetanzSophie Madeleine NagelJulian KünzelMadita WandreyJan HagemannRoland H. StauberChristian FreeseDésirée GülMDPI AGarticleautomationliquid biopsycirculating tumor cellshead and neck squamous cell carcinomaimmunomagnetic particle-based detectionmetastasisMedicine (General)R5-920ENDiagnostics, Vol 11, Iss 2040, p 2040 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	automation liquid biopsy circulating tumor cells head and neck squamous cell carcinoma immunomagnetic particle-based detection metastasis Medicine (General) R5-920
spellingShingle	automation liquid biopsy circulating tumor cells head and neck squamous cell carcinoma immunomagnetic particle-based detection metastasis Medicine (General) R5-920 Alena Gribko Janis Stiefel Lana Liebetanz Sophie Madeleine Nagel Julian Künzel Madita Wandrey Jan Hagemann Roland H. Stauber Christian Freese Désirée Gül IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells
description	Background: detailed information about circulating tumor cells (CTCs) as an indicator of therapy response and cancer metastasis is crucial not only for basic research but also for diagnostics and therapeutic approaches. Here, we showcase a newly developed IsoMAG IMS system with an optimized protocol for fully automated immunomagnetic enrichment of CTCs, also revealing rare CTC subpopulations. Methods: using different squamous cell carcinoma cell lines, we developed an isolation protocol exploiting highly efficient EpCAM-targeting magnetic beads for automated CTC enrichment by the IsoMAG IMS system. By FACS analysis, we analyzed white blood contamination usually preventing further downstream analysis of enriched cells. Results: 1 µm magnetic beads with tosyl-activated hydrophobic surface properties were found to be optimal for automated CTC enrichment. More than 86.5% and 95% of spiked cancer cells were recovered from both cell culture media or human blood employing our developed protocol. In addition, contamination with white blood cells was minimized to about 1200 cells starting from 7.5 mL blood. Finally, we showed that the system is applicable for HNSCC patient samples and characterized isolated CTCs by immunostaining using a panel of tumor markers. Conclusion: Herein, we demonstrate that the IsoMAG system allows the detection and isolation of CTCs from HNSCC patient blood for disease monitoring in a fully-automated process with a significant leukocyte count reduction. Future developments seek to integrate the IsoMAG IMS system into an automated microfluidic-based isolation workflow to further facilitate single CTC detection also in clinical routine.
format	article
author	Alena Gribko Janis Stiefel Lana Liebetanz Sophie Madeleine Nagel Julian Künzel Madita Wandrey Jan Hagemann Roland H. Stauber Christian Freese Désirée Gül
author_facet	Alena Gribko Janis Stiefel Lana Liebetanz Sophie Madeleine Nagel Julian Künzel Madita Wandrey Jan Hagemann Roland H. Stauber Christian Freese Désirée Gül
author_sort	Alena Gribko
title	IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells
title_short	IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells
title_full	IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells
title_fullStr	IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells
title_full_unstemmed	IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells
title_sort	isomag—an automated system for the immunomagnetic isolation of squamous cell carcinoma-derived circulating tumor cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/530d9fbd81f045eba42d343f7ae680f5
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IsoMAG—An Automated System for the Immunomagnetic Isolation of Squamous Cell Carcinoma-Derived Circulating Tumor Cells

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