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CTC enrichment and sorting

FG Francesca Galardi
FL Francesca De Luca
CB Chiara Biagioni
IM Ilenia Migliaccio
GC Giuseppe Curigliano
AM Alessandro M. Minisini
MB Martina Bonechi
EM Erica Moretti
ER Emanuela Risi
AM Amelia McCartney
MB Matteo Benelli
DR Dario Romagnoli
SC Silvia Cappadona
SG Stefano Gabellini
CG Cristina Guarducci
VC Valerio Conti
LB Laura Biganzoli
AL Angelo Di Leo
LM Luca Malorni
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CTC enrichment and enumeration were carried out by CellSearch® System (CS) (Menarini Silicon Biosystems). CTCs were isolated from 7.5 mL of whole blood with CellSearch™Epithelial Cell kit. After enrichment, isolated and stained cells were identified and counted in the CellTracks® Analyzer II. Cartridges from patient samples with five or more CTCs were kept in the dark at 4 °C before cell sorting by DEPAarray™ System (Menarini Silicon Biosystems).

For DEPAarray sorting, stained cells in suspension were recovered from CellSearch cartridges and, after a volume reduction, were loaded into the DA A300K DS V2.0 chip (Menarini Silicon Biosystems) and set into the DA system, according to the manufacturer’s instructions. After identification, cells were manipulated by dielectrophoresis cages and recovered as single CTCs or pools of pure CTCs into 200 μL tubes. CTC samples were then stored at − 20 °C for later RNA extraction, according to the manufacturer’s protocol.

Copyright and License information: The Author(s) ©2021
Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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