Potential use of fish collagen matrix to evaluate tubulogenesis in vitro

Autores
Medina, Daiana Mailen; Acevedo Gomez, Antonella Valeria; Leiva, Laura Cristina Ana; Pellegrini Malpiedi, Luciana; Bustillo, Soledad
Año de publicación
2022
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Formation or sprouting of new blood vessels (angiogenesis) is a complex process that involves the extracellular matrix (ECM) and endothelial cells (EC). A common in vitro angiogenesis assay consists of culturing EC on or inside distinct ECM components such as collagen. Recently, collagen from aquatic sources has gained attention in tissue engineering applications. Thus, in this work the potential of fish collagen from Pygocentrus nattereri (CP) to induce tubular structures in tEnd.1 (RRID: CVCL_6272) cell line was evaluated. Firstly, collagen from discarded fish skin was extracted by acid solubilization, concentrated by salt precipitation, dialyzed against 100 mM acid acetic and preserve at 8°C. The hydroxyproline analysis revealed a collagen concentration of 6,9 mg/mL (73,8 % of total protein content) and the electrophoretic pattern (SDS-PAGE under non-reducing conditions) showed that extracted CP was type I. To evaluate CP use in angiogenesis assay, 100 ⎧L of a 0.5 mg/mL solution (culture medium-pH 7) was added to each well of 96-well plate and incubated for 30 min at 37°C. Endothelial cells (30.000 cells/well DMEM-5%FBS) were seeded on collagen-coated wells and incubated for 24h at 37°C-5%-CO2. Controls were performed using untreated wells. Tube formation was monitored by an inverted phase contrast microscope and images were taken with a digital camera. Using ImageJ software, number of capillary structures was analysed. EC cultured in collagen matrixes organized rapidly into an irregular network, which was formed by many tubular structures radiating out from cell aggregates. These results suggest the potential use of fish collagen in cell culture.
Fil: Medina, Daiana Mailen. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Acevedo Gomez, Antonella Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
Fil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Bustillo, Soledad. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
IX Simposio Latinoamericano de Tecnología de Cultivos Celulares
Santa Fe
Argentina
Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Centro Biotecnológico del Litoral
Materia
COLLAGEN
TUBULOGENESIS
FISH
MATRIX
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/203906
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Potential use of fish collagen matrix to evaluate tubulogenesis in vitroMedina, Daiana MailenAcevedo Gomez, Antonella ValeriaLeiva, Laura Cristina AnaPellegrini Malpiedi, LucianaBustillo, SoledadCOLLAGENTUBULOGENESISFISHMATRIXhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Formation or sprouting of new blood vessels (angiogenesis) is a complex process that involves the extracellular matrix (ECM) and endothelial cells (EC). A common in vitro angiogenesis assay consists of culturing EC on or inside distinct ECM components such as collagen. Recently, collagen from aquatic sources has gained attention in tissue engineering applications. Thus, in this work the potential of fish collagen from Pygocentrus nattereri (CP) to induce tubular structures in tEnd.1 (RRID: CVCL_6272) cell line was evaluated. Firstly, collagen from discarded fish skin was extracted by acid solubilization, concentrated by salt precipitation, dialyzed against 100 mM acid acetic and preserve at 8°C. The hydroxyproline analysis revealed a collagen concentration of 6,9 mg/mL (73,8 % of total protein content) and the electrophoretic pattern (SDS-PAGE under non-reducing conditions) showed that extracted CP was type I. To evaluate CP use in angiogenesis assay, 100 ⎧L of a 0.5 mg/mL solution (culture medium-pH 7) was added to each well of 96-well plate and incubated for 30 min at 37°C. Endothelial cells (30.000 cells/well DMEM-5%FBS) were seeded on collagen-coated wells and incubated for 24h at 37°C-5%-CO2. Controls were performed using untreated wells. Tube formation was monitored by an inverted phase contrast microscope and images were taken with a digital camera. Using ImageJ software, number of capillary structures was analysed. EC cultured in collagen matrixes organized rapidly into an irregular network, which was formed by many tubular structures radiating out from cell aggregates. These results suggest the potential use of fish collagen in cell culture.Fil: Medina, Daiana Mailen. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Acevedo Gomez, Antonella Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaFil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Bustillo, Soledad. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaIX Simposio Latinoamericano de Tecnología de Cultivos CelularesSanta FeArgentinaUniversidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Centro Biotecnológico del LitoralUniversidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Centro Biotecnológico del Litoral2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectSimposioBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/203906Potential use of fish collagen matrix to evaluate tubulogenesis in vitro; IX Simposio Latinoamericano de Tecnología de Cultivos Celulares; Santa Fe; Argentina; 2022; 42-42CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.fbcb.unl.edu.ar/slatcc2022/wp-content/uploads/sites/12/2020/02/Libro-SLATCC-version-virtual_221027.pdfinfo:eu-repo/semantics/altIdentifier/url/https://www.fbcb.unl.edu.ar/slatcc2022/Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-03T09:19:42Zoai:ri.conicet.gov.ar:11336/203906instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-12-03 09:19:42.646CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
title Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
spellingShingle Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
Medina, Daiana Mailen
COLLAGEN
TUBULOGENESIS
FISH
MATRIX
title_short Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
title_full Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
title_fullStr Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
title_full_unstemmed Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
title_sort Potential use of fish collagen matrix to evaluate tubulogenesis in vitro
dc.creator.none.fl_str_mv Medina, Daiana Mailen
Acevedo Gomez, Antonella Valeria
Leiva, Laura Cristina Ana
Pellegrini Malpiedi, Luciana
Bustillo, Soledad
author Medina, Daiana Mailen
author_facet Medina, Daiana Mailen
Acevedo Gomez, Antonella Valeria
Leiva, Laura Cristina Ana
Pellegrini Malpiedi, Luciana
Bustillo, Soledad
author_role author
author2 Acevedo Gomez, Antonella Valeria
Leiva, Laura Cristina Ana
Pellegrini Malpiedi, Luciana
Bustillo, Soledad
author2_role author
author
author
author
dc.subject.none.fl_str_mv COLLAGEN
TUBULOGENESIS
FISH
MATRIX
topic COLLAGEN
TUBULOGENESIS
FISH
MATRIX
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Formation or sprouting of new blood vessels (angiogenesis) is a complex process that involves the extracellular matrix (ECM) and endothelial cells (EC). A common in vitro angiogenesis assay consists of culturing EC on or inside distinct ECM components such as collagen. Recently, collagen from aquatic sources has gained attention in tissue engineering applications. Thus, in this work the potential of fish collagen from Pygocentrus nattereri (CP) to induce tubular structures in tEnd.1 (RRID: CVCL_6272) cell line was evaluated. Firstly, collagen from discarded fish skin was extracted by acid solubilization, concentrated by salt precipitation, dialyzed against 100 mM acid acetic and preserve at 8°C. The hydroxyproline analysis revealed a collagen concentration of 6,9 mg/mL (73,8 % of total protein content) and the electrophoretic pattern (SDS-PAGE under non-reducing conditions) showed that extracted CP was type I. To evaluate CP use in angiogenesis assay, 100 ⎧L of a 0.5 mg/mL solution (culture medium-pH 7) was added to each well of 96-well plate and incubated for 30 min at 37°C. Endothelial cells (30.000 cells/well DMEM-5%FBS) were seeded on collagen-coated wells and incubated for 24h at 37°C-5%-CO2. Controls were performed using untreated wells. Tube formation was monitored by an inverted phase contrast microscope and images were taken with a digital camera. Using ImageJ software, number of capillary structures was analysed. EC cultured in collagen matrixes organized rapidly into an irregular network, which was formed by many tubular structures radiating out from cell aggregates. These results suggest the potential use of fish collagen in cell culture.
Fil: Medina, Daiana Mailen. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Acevedo Gomez, Antonella Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
Fil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Bustillo, Soledad. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
IX Simposio Latinoamericano de Tecnología de Cultivos Celulares
Santa Fe
Argentina
Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Centro Biotecnológico del Litoral
description Formation or sprouting of new blood vessels (angiogenesis) is a complex process that involves the extracellular matrix (ECM) and endothelial cells (EC). A common in vitro angiogenesis assay consists of culturing EC on or inside distinct ECM components such as collagen. Recently, collagen from aquatic sources has gained attention in tissue engineering applications. Thus, in this work the potential of fish collagen from Pygocentrus nattereri (CP) to induce tubular structures in tEnd.1 (RRID: CVCL_6272) cell line was evaluated. Firstly, collagen from discarded fish skin was extracted by acid solubilization, concentrated by salt precipitation, dialyzed against 100 mM acid acetic and preserve at 8°C. The hydroxyproline analysis revealed a collagen concentration of 6,9 mg/mL (73,8 % of total protein content) and the electrophoretic pattern (SDS-PAGE under non-reducing conditions) showed that extracted CP was type I. To evaluate CP use in angiogenesis assay, 100 ⎧L of a 0.5 mg/mL solution (culture medium-pH 7) was added to each well of 96-well plate and incubated for 30 min at 37°C. Endothelial cells (30.000 cells/well DMEM-5%FBS) were seeded on collagen-coated wells and incubated for 24h at 37°C-5%-CO2. Controls were performed using untreated wells. Tube formation was monitored by an inverted phase contrast microscope and images were taken with a digital camera. Using ImageJ software, number of capillary structures was analysed. EC cultured in collagen matrixes organized rapidly into an irregular network, which was formed by many tubular structures radiating out from cell aggregates. These results suggest the potential use of fish collagen in cell culture.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
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Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/203906
Potential use of fish collagen matrix to evaluate tubulogenesis in vitro; IX Simposio Latinoamericano de Tecnología de Cultivos Celulares; Santa Fe; Argentina; 2022; 42-42
CONICET Digital
CONICET
url http://hdl.handle.net/11336/203906
identifier_str_mv Potential use of fish collagen matrix to evaluate tubulogenesis in vitro; IX Simposio Latinoamericano de Tecnología de Cultivos Celulares; Santa Fe; Argentina; 2022; 42-42
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.fbcb.unl.edu.ar/slatcc2022/wp-content/uploads/sites/12/2020/02/Libro-SLATCC-version-virtual_221027.pdf
info:eu-repo/semantics/altIdentifier/url/https://www.fbcb.unl.edu.ar/slatcc2022/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Centro Biotecnológico del Litoral
publisher.none.fl_str_mv Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Centro Biotecnológico del Litoral
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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