Cargando…

Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients

Detailed examination of tumor components is leading‐edge to establish personalized cancer therapy. Accompanying research on cell‐free DNA, the cell count of circulating tumor cells (CTCs) in patient blood is seen as a crucial prognostic factor. The potential of CTC analysis is further not limited to...

Descripción completa

Detalles Bibliográficos
Autores principales: Stiefel, Janis, Freese, Christian, Sriram, Ashwin, Alebrand, Sabine, Srinivas, Nalini, Sproll, Christoph, Wandrey, Madita, Gül, Désirée, Hagemann, Jan, Becker, Jürgen C., Baßler, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9077830/
https://www.ncbi.nlm.nih.gov/pubmed/35573135
http://dx.doi.org/10.1002/elsc.202100133
_version_ 1784702197527740416
author Stiefel, Janis
Freese, Christian
Sriram, Ashwin
Alebrand, Sabine
Srinivas, Nalini
Sproll, Christoph
Wandrey, Madita
Gül, Désirée
Hagemann, Jan
Becker, Jürgen C.
Baßler, Michael
author_facet Stiefel, Janis
Freese, Christian
Sriram, Ashwin
Alebrand, Sabine
Srinivas, Nalini
Sproll, Christoph
Wandrey, Madita
Gül, Désirée
Hagemann, Jan
Becker, Jürgen C.
Baßler, Michael
author_sort Stiefel, Janis
collection PubMed
description Detailed examination of tumor components is leading‐edge to establish personalized cancer therapy. Accompanying research on cell‐free DNA, the cell count of circulating tumor cells (CTCs) in patient blood is seen as a crucial prognostic factor. The potential of CTC analysis is further not limited to the determination of the overall survival rate but sheds light on understanding inter‐ and intratumoral heterogeneity. In this regard, commercial CTC isolation devices combining an efficient enrichment of rare cells with a droplet deposition of single cells for downstream analysis are highly appreciated. The Liquid biopsy platform CTCelect was developed to realize a fully‐automated enrichment and single cell dispensing of CTCs from whole blood without pre‐processing. We characterized each process step with two different carcinoma cell lines demonstrating up to 87 % enrichment (n = 10) with EpCAM coupled immunomagnetic beads, 73 % optical detection and dispensing efficiency (n = 5). 40 to 56.7 % of cells were recovered after complete isolation from 7.5 ml untreated whole blood (n = 6). In this study, CTCelect enabled automated dispensing of single circulating tumor cells from HNSCC patient samples, qPCR‐based confirmation of tumor‐related biomarkers and immunostaining. Finally, the platform was compared to commercial CTC isolation technologies to highlight advantages and limitations of CTCelect. This system offers new possibilities for single cell screening in cancer diagnostics, individual therapy approaches and real‐time monitoring.
format Online
Article
Text
id pubmed-9077830
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-90778302022-05-13 Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients Stiefel, Janis Freese, Christian Sriram, Ashwin Alebrand, Sabine Srinivas, Nalini Sproll, Christoph Wandrey, Madita Gül, Désirée Hagemann, Jan Becker, Jürgen C. Baßler, Michael Eng Life Sci Research Articles Detailed examination of tumor components is leading‐edge to establish personalized cancer therapy. Accompanying research on cell‐free DNA, the cell count of circulating tumor cells (CTCs) in patient blood is seen as a crucial prognostic factor. The potential of CTC analysis is further not limited to the determination of the overall survival rate but sheds light on understanding inter‐ and intratumoral heterogeneity. In this regard, commercial CTC isolation devices combining an efficient enrichment of rare cells with a droplet deposition of single cells for downstream analysis are highly appreciated. The Liquid biopsy platform CTCelect was developed to realize a fully‐automated enrichment and single cell dispensing of CTCs from whole blood without pre‐processing. We characterized each process step with two different carcinoma cell lines demonstrating up to 87 % enrichment (n = 10) with EpCAM coupled immunomagnetic beads, 73 % optical detection and dispensing efficiency (n = 5). 40 to 56.7 % of cells were recovered after complete isolation from 7.5 ml untreated whole blood (n = 6). In this study, CTCelect enabled automated dispensing of single circulating tumor cells from HNSCC patient samples, qPCR‐based confirmation of tumor‐related biomarkers and immunostaining. Finally, the platform was compared to commercial CTC isolation technologies to highlight advantages and limitations of CTCelect. This system offers new possibilities for single cell screening in cancer diagnostics, individual therapy approaches and real‐time monitoring. John Wiley and Sons Inc. 2022-04-06 /pmc/articles/PMC9077830/ /pubmed/35573135 http://dx.doi.org/10.1002/elsc.202100133 Text en © 2022 The Authors. Engineering in Life Sciences published by Wiley‐VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Stiefel, Janis
Freese, Christian
Sriram, Ashwin
Alebrand, Sabine
Srinivas, Nalini
Sproll, Christoph
Wandrey, Madita
Gül, Désirée
Hagemann, Jan
Becker, Jürgen C.
Baßler, Michael
Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients
title Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients
title_full Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients
title_fullStr Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients
title_full_unstemmed Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients
title_short Characterization of a novel microfluidic platform for the isolation of rare single cells to enable CTC analysis from head and neck squamous cell carcinoma patients
title_sort characterization of a novel microfluidic platform for the isolation of rare single cells to enable ctc analysis from head and neck squamous cell carcinoma patients
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9077830/
https://www.ncbi.nlm.nih.gov/pubmed/35573135
http://dx.doi.org/10.1002/elsc.202100133
work_keys_str_mv AT stiefeljanis characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT freesechristian characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT sriramashwin characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT alebrandsabine characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT srinivasnalini characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT sprollchristoph characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT wandreymadita characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT guldesiree characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT hagemannjan characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT beckerjurgenc characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients
AT baßlermichael characterizationofanovelmicrofluidicplatformfortheisolationofraresinglecellstoenablectcanalysisfromheadandnecksquamouscellcarcinomapatients