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Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules

In the present report, we are making the proof of concept of cell small populations (from 1 to 100 cells) spotting, culture and secretion detection on a gold surface. In order to keep the cells in a hydrated environment during the robotized micropipetting and to address different cell lines on a sin...

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Autores principales: Berthuy, Ophélie I., Blum, Loïc J., Marquette, Christophe A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4810394/
https://www.ncbi.nlm.nih.gov/pubmed/26784243
http://dx.doi.org/10.3390/bios6010002
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author Berthuy, Ophélie I.
Blum, Loïc J.
Marquette, Christophe A.
author_facet Berthuy, Ophélie I.
Blum, Loïc J.
Marquette, Christophe A.
author_sort Berthuy, Ophélie I.
collection PubMed
description In the present report, we are making the proof of concept of cell small populations (from 1 to 100 cells) spotting, culture and secretion detection on a gold surface. In order to keep the cells in a hydrated environment during the robotized micropipetting and to address different cell lines on a single chip, a biocompatible alginate polymer was used. This approach enables the encapsulation of the cell in a very small volume (30 nL), directly on the substrate and permits a precise control of the number of cells in each alginate bead. After 24 h of culture, the adherent cells are ready for surface plasmon resonance imaging (SPRi) experimentation. To enable the detection of secreted proteins, various antibodies are immobilized in an organized manner on a SPRi sensor and permitted the multiplex detection of different proteins secreted by the different cultured cell lines. Evidence of the real-time detection will be presented for Prostate Specific Antigen (PSA) and β-2-microglobulin (B2M) secreted by prostate cancer cells following induction by dihydrotestosterone (DHT). Different kinetics for the two secreted proteins were then demonstrated and precisely determined using the chip.
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spelling pubmed-48103942016-04-04 Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules Berthuy, Ophélie I. Blum, Loïc J. Marquette, Christophe A. Biosensors (Basel) Article In the present report, we are making the proof of concept of cell small populations (from 1 to 100 cells) spotting, culture and secretion detection on a gold surface. In order to keep the cells in a hydrated environment during the robotized micropipetting and to address different cell lines on a single chip, a biocompatible alginate polymer was used. This approach enables the encapsulation of the cell in a very small volume (30 nL), directly on the substrate and permits a precise control of the number of cells in each alginate bead. After 24 h of culture, the adherent cells are ready for surface plasmon resonance imaging (SPRi) experimentation. To enable the detection of secreted proteins, various antibodies are immobilized in an organized manner on a SPRi sensor and permitted the multiplex detection of different proteins secreted by the different cultured cell lines. Evidence of the real-time detection will be presented for Prostate Specific Antigen (PSA) and β-2-microglobulin (B2M) secreted by prostate cancer cells following induction by dihydrotestosterone (DHT). Different kinetics for the two secreted proteins were then demonstrated and precisely determined using the chip. MDPI 2016-01-15 /pmc/articles/PMC4810394/ /pubmed/26784243 http://dx.doi.org/10.3390/bios6010002 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Berthuy, Ophélie I.
Blum, Loïc J.
Marquette, Christophe A.
Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules
title Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules
title_full Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules
title_fullStr Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules
title_full_unstemmed Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules
title_short Cancer-Cells on Chip for Label-Free Detection of Secreted Molecules
title_sort cancer-cells on chip for label-free detection of secreted molecules
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4810394/
https://www.ncbi.nlm.nih.gov/pubmed/26784243
http://dx.doi.org/10.3390/bios6010002
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