Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates

The effect of cell-cell contact on gene transfection is mainly unknown. Usually, transfection is carried out in batch cell cultures without control over cellular interactions, and efficiency analysis relies on complex and expensive protocols commonly involving flow cytometry as the final analytical...

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Autores: Azuaje Hualde, Enrique, Rosique, Melania, Calatayud Sánchez, Alba, Benito López, Fernando, Basabe Desmonts, Lourdes, Martínez de Pancorbo Gómez, María de los Angeles
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universidad del País Vasco
Repositorio:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/51055
Acceso en línea:http://hdl.handle.net/10810/51055
Access Level:acceso abierto
Palabra clave:surface patterning
cell patterning
gene transfection
cell-cellcontact
continuous monitoring
GFP
mesenchymal stromalcell
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spelling Continuous Monitoring of Cell Transfection Efficiency with Micropatterned SubstratesAzuaje Hualde, EnriqueRosique, MelaniaCalatayud Sánchez, AlbaBenito López, FernandoBasabe Desmonts, LourdesMartínez de Pancorbo Gómez, María de los Angelessurface patterningcell patterninggene transfectioncell-cellcontactcontinuous monitoringGFPmesenchymal stromalcellThe effect of cell-cell contact on gene transfection is mainly unknown. Usually, transfection is carried out in batch cell cultures without control over cellular interactions, and efficiency analysis relies on complex and expensive protocols commonly involving flow cytometry as the final analytical step. Novel platforms and cell patterning are being studied in order to control cellular interactions and improve quantification methods. In this work, we report the use of surface patterning of fibronectin for the generation of two types of mesenchymal stromal cells patterns: single cell patterns without cell-to-cell contact, and small cell-colonypatterns. Both scenarios allowed the integration of the full transfection process and the continuous monitoring of thousands of individualized events by fluorescence microscopy. Our results showed that cell-to-cell contact clearly affected the transfection, as single cells presented a maximum transfection peak 6 hours earlier and had a 10 % higher transfection efficiency than cells with cell-to-cell contact.Authors acknowledge funding support from Basque Government, under Grupos Consolidados with Grant No. IT1271-19 and from Gobierno de España, Ministerio de Economia y Competitividad, with Grant No. BIO2016-80417-P(AEI/FEDER, UE). This project has received funding from the European Union, DNASurf Horizon2020 project [#778001],funded through the Marie Sklodowska-Curie Actions RISE programme for international and intersectoral Research and Innovation Staff Exchange. E.A.-H. acknowledges funding from the BasqueGovernment, Department of Education, for predoctoral fellowship 2016. Authors thank Dr. Maite Alvarez fromtheDNA Bank Service (SGIker) of the University of the Basque Country (UPV/EHU) and European funding (ERDF and ESF)for technical and human support.Authors thank the intellectual and technical assistance from the ICTS “NANBIOSIS”, more specifically by the Drug Formulation Unit (U10) of the CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN).Authors thank Dr. Alberto Gorrochategui from “Clínica Dermatológica Ercilla (Bilbao)” for providing the hair follicles from where hHF-MSCs were extractedWileyEuropean Commission202120212021info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/51055reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoInglésinfo:eu-repo/grantAgreement/EC/H2020/778001info:eu-repo/grantAgreement/MINECO/BIO2016-80417-P/https://doi.org/10.1002/bit.27783info:eu-repo/semantics/openAccess(c) 2021 Wileyoai:addi.ehu.eus:10810/510552026-06-18T09:23:17Z
dc.title.none.fl_str_mv Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
title Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
spellingShingle Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
Azuaje Hualde, Enrique
surface patterning
cell patterning
gene transfection
cell-cellcontact
continuous monitoring
GFP
mesenchymal stromalcell
title_short Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
title_full Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
title_fullStr Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
title_full_unstemmed Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
title_sort Continuous Monitoring of Cell Transfection Efficiency with Micropatterned Substrates
dc.creator.none.fl_str_mv Azuaje Hualde, Enrique
Rosique, Melania
Calatayud Sánchez, Alba
Benito López, Fernando
Basabe Desmonts, Lourdes
Martínez de Pancorbo Gómez, María de los Angeles
author Azuaje Hualde, Enrique
author_facet Azuaje Hualde, Enrique
Rosique, Melania
Calatayud Sánchez, Alba
Benito López, Fernando
Basabe Desmonts, Lourdes
Martínez de Pancorbo Gómez, María de los Angeles
author_role author
author2 Rosique, Melania
Calatayud Sánchez, Alba
Benito López, Fernando
Basabe Desmonts, Lourdes
Martínez de Pancorbo Gómez, María de los Angeles
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv European Commission
dc.subject.none.fl_str_mv surface patterning
cell patterning
gene transfection
cell-cellcontact
continuous monitoring
GFP
mesenchymal stromalcell
topic surface patterning
cell patterning
gene transfection
cell-cellcontact
continuous monitoring
GFP
mesenchymal stromalcell
description The effect of cell-cell contact on gene transfection is mainly unknown. Usually, transfection is carried out in batch cell cultures without control over cellular interactions, and efficiency analysis relies on complex and expensive protocols commonly involving flow cytometry as the final analytical step. Novel platforms and cell patterning are being studied in order to control cellular interactions and improve quantification methods. In this work, we report the use of surface patterning of fibronectin for the generation of two types of mesenchymal stromal cells patterns: single cell patterns without cell-to-cell contact, and small cell-colonypatterns. Both scenarios allowed the integration of the full transfection process and the continuous monitoring of thousands of individualized events by fluorescence microscopy. Our results showed that cell-to-cell contact clearly affected the transfection, as single cells presented a maximum transfection peak 6 hours earlier and had a 10 % higher transfection efficiency than cells with cell-to-cell contact.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/51055
url http://hdl.handle.net/10810/51055
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/EC/H2020/778001
info:eu-repo/grantAgreement/MINECO/BIO2016-80417-P/
https://doi.org/10.1002/bit.27783
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
(c) 2021 Wiley
eu_rights_str_mv openAccess
rights_invalid_str_mv (c) 2021 Wiley
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869413255130120192
score 15.301629

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