Genetically encoded sender-receiver system in 3D mammalian cell culture

Autores
Carvalho, A.; Bárcena Menéndez, Diego; Senthivel, V. R.; Zimmermann, T.; Diambra, Luis Aníbal; Isalan, M.
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Engineering spatial patterning in mammalian cells, employing entirely genetically encoded components, requires solving several problems. These include how to code secreted activator or inhibitor molecules and how to send concentration-dependent signals to neighboring cells, to control gene expression. The Madin-Darby Canine Kidney (MDCK) cell line is a potential engineering scaffold as it forms hollow spheres (cysts) in 3D culture and tubulates in response to extracellular hepatocyte growth factor (HGF). We first aimed to graft a synthetic patterning system onto single developing MDCK cysts. We therefore developed a new localized transfection method to engineer distinct sender and receiver regions. A stable reporter line enabled reversible EGFP activation by HGF and modulation by a secreted repressor (a truncated HGF variant, NK4). By expanding the scale to wide fields of cysts, we generated morphogen diffusion gradients, controlling reporter gene expression. Together, these components provide a toolkit for engineering cell-cell communication networks in 3D cell culture.
Centro Regional de Estudios Genómicos
Materia
Ciencias Exactas
HGF
MDCK
morphogen
NK4
synthetic patterning
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/85585
Acceder
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oai_identifier_str oai:sedici.unlp.edu.ar:10915/85585
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Genetically encoded sender-receiver system in 3D mammalian cell cultureCarvalho, A.Bárcena Menéndez, DiegoSenthivel, V. R.Zimmermann, T.Diambra, Luis AníbalIsalan, M.Ciencias ExactasHGFMDCKmorphogenNK4synthetic patterningEngineering spatial patterning in mammalian cells, employing entirely genetically encoded components, requires solving several problems. These include how to code secreted activator or inhibitor molecules and how to send concentration-dependent signals to neighboring cells, to control gene expression. The Madin-Darby Canine Kidney (MDCK) cell line is a potential engineering scaffold as it forms hollow spheres (cysts) in 3D culture and tubulates in response to extracellular hepatocyte growth factor (HGF). We first aimed to graft a synthetic patterning system onto single developing MDCK cysts. We therefore developed a new localized transfection method to engineer distinct sender and receiver regions. A stable reporter line enabled reversible EGFP activation by HGF and modulation by a secreted repressor (a truncated HGF variant, NK4). By expanding the scale to wide fields of cysts, we generated morphogen diffusion gradients, controlling reporter gene expression. Together, these components provide a toolkit for engineering cell-cell communication networks in 3D cell culture.Centro Regional de Estudios Genómicos2014info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf264-272http://sedici.unlp.edu.ar/handle/10915/85585enginfo:eu-repo/semantics/altIdentifier/issn/2161-5063info:eu-repo/semantics/altIdentifier/doi/10.1021/sb400053binfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2026-01-07T13:00:25Zoai:sedici.unlp.edu.ar:10915/85585Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292026-01-07 13:00:25.323SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Genetically encoded sender-receiver system in 3D mammalian cell culture
title Genetically encoded sender-receiver system in 3D mammalian cell culture
spellingShingle Genetically encoded sender-receiver system in 3D mammalian cell culture
Carvalho, A.
Ciencias Exactas
HGF
MDCK
morphogen
NK4
synthetic patterning
title_short Genetically encoded sender-receiver system in 3D mammalian cell culture
title_full Genetically encoded sender-receiver system in 3D mammalian cell culture
title_fullStr Genetically encoded sender-receiver system in 3D mammalian cell culture
title_full_unstemmed Genetically encoded sender-receiver system in 3D mammalian cell culture
title_sort Genetically encoded sender-receiver system in 3D mammalian cell culture
dc.creator.none.fl_str_mv Carvalho, A.
Bárcena Menéndez, Diego
Senthivel, V. R.
Zimmermann, T.
Diambra, Luis Aníbal
Isalan, M.
author Carvalho, A.
author_facet Carvalho, A.
Bárcena Menéndez, Diego
Senthivel, V. R.
Zimmermann, T.
Diambra, Luis Aníbal
Isalan, M.
author_role author
author2 Bárcena Menéndez, Diego
Senthivel, V. R.
Zimmermann, T.
Diambra, Luis Aníbal
Isalan, M.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Ciencias Exactas
HGF
MDCK
morphogen
NK4
synthetic patterning
topic Ciencias Exactas
HGF
MDCK
morphogen
NK4
synthetic patterning
dc.description.none.fl_txt_mv Engineering spatial patterning in mammalian cells, employing entirely genetically encoded components, requires solving several problems. These include how to code secreted activator or inhibitor molecules and how to send concentration-dependent signals to neighboring cells, to control gene expression. The Madin-Darby Canine Kidney (MDCK) cell line is a potential engineering scaffold as it forms hollow spheres (cysts) in 3D culture and tubulates in response to extracellular hepatocyte growth factor (HGF). We first aimed to graft a synthetic patterning system onto single developing MDCK cysts. We therefore developed a new localized transfection method to engineer distinct sender and receiver regions. A stable reporter line enabled reversible EGFP activation by HGF and modulation by a secreted repressor (a truncated HGF variant, NK4). By expanding the scale to wide fields of cysts, we generated morphogen diffusion gradients, controlling reporter gene expression. Together, these components provide a toolkit for engineering cell-cell communication networks in 3D cell culture.
Centro Regional de Estudios Genómicos
description Engineering spatial patterning in mammalian cells, employing entirely genetically encoded components, requires solving several problems. These include how to code secreted activator or inhibitor molecules and how to send concentration-dependent signals to neighboring cells, to control gene expression. The Madin-Darby Canine Kidney (MDCK) cell line is a potential engineering scaffold as it forms hollow spheres (cysts) in 3D culture and tubulates in response to extracellular hepatocyte growth factor (HGF). We first aimed to graft a synthetic patterning system onto single developing MDCK cysts. We therefore developed a new localized transfection method to engineer distinct sender and receiver regions. A stable reporter line enabled reversible EGFP activation by HGF and modulation by a secreted repressor (a truncated HGF variant, NK4). By expanding the scale to wide fields of cysts, we generated morphogen diffusion gradients, controlling reporter gene expression. Together, these components provide a toolkit for engineering cell-cell communication networks in 3D cell culture.
publishDate 2014
dc.date.none.fl_str_mv 2014
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://sedici.unlp.edu.ar/handle/10915/85585
url http://sedici.unlp.edu.ar/handle/10915/85585
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/2161-5063
info:eu-repo/semantics/altIdentifier/doi/10.1021/sb400053b
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
264-272
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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score 13.25844

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