Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels

Detalhes bibliográficos
Autor(a) principal: Cairrão, Elisa
Data de Publicação: 2009
Outros Autores: Santos-Silva, António, Alvarez, Ezequiel, Correia, Ilídio Joaquim Sobreira, Verde, Ignacio
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.6/4622
Resumo: The human umbilical cord is a biological sample that can be easily obtained just after birth. A methodology was developed to perform cultures of human umbilical artery smooth muscle cells (HUASMC) expressing contractile proteins and functional ionic channels. To avoid fibroblast and endothelial cell contamination, we mechanically separated the tunica media, which only contains HUASMC and matrix proteins. To isolate the cells, collagenase V and elastase were used as hydrolyzing enzymes. The isolated cells were plated in collagen-coated dishes to obtain cultures of HUASMC. The cells obtained after different passages (1 to 6) exhibit the characteristic vascular smooth cell morphology and express smooth muscle alpha-2 actin, myosin heavy chain SM1, and alpha subunits of L- and T-type calcium channels (Cav 1.2, Cav 1.2, and Cav 3.2). Electrophysiology recordings for L- and T-type calcium channels were made, indicating that these channels are functional in the cultured cells. In conclusion, the procedure developed allows obtaining cultures of HUASMC expressing contractile proteins and also functional ionic channels. These cells could be used to study cellular and molecular aspects about the regulation of the vascular function.
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spelling Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channelsUmbilical arterySmooth muscle cellsPatch-clampCalcium channelsThe human umbilical cord is a biological sample that can be easily obtained just after birth. A methodology was developed to perform cultures of human umbilical artery smooth muscle cells (HUASMC) expressing contractile proteins and functional ionic channels. To avoid fibroblast and endothelial cell contamination, we mechanically separated the tunica media, which only contains HUASMC and matrix proteins. To isolate the cells, collagenase V and elastase were used as hydrolyzing enzymes. The isolated cells were plated in collagen-coated dishes to obtain cultures of HUASMC. The cells obtained after different passages (1 to 6) exhibit the characteristic vascular smooth cell morphology and express smooth muscle alpha-2 actin, myosin heavy chain SM1, and alpha subunits of L- and T-type calcium channels (Cav 1.2, Cav 1.2, and Cav 3.2). Electrophysiology recordings for L- and T-type calcium channels were made, indicating that these channels are functional in the cultured cells. In conclusion, the procedure developed allows obtaining cultures of HUASMC expressing contractile proteins and also functional ionic channels. These cells could be used to study cellular and molecular aspects about the regulation of the vascular function.SpringeruBibliorumCairrão, ElisaSantos-Silva, AntónioAlvarez, EzequielCorreia, Ilídio Joaquim SobreiraVerde, Ignacio2018-03-15T16:00:29Z2009-01-012009-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.6/4622engCairrão, E., Santos-Silva, A.J., Alvarez, E., Correia, I.J. e Verde, I. (2009) “Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels”, In Vitro Cellular & Developmental Biology – Animal, Vol. 45 (3), pp. 175-18410.1007/s11626-008-9161-6metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-12-15T09:41:41Zoai:ubibliorum.ubi.pt:10400.6/4622Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:45:41.168648Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
title Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
spellingShingle Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
Cairrão, Elisa
Umbilical artery
Smooth muscle cells
Patch-clamp
Calcium channels
title_short Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
title_full Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
title_fullStr Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
title_full_unstemmed Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
title_sort Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
author Cairrão, Elisa
author_facet Cairrão, Elisa
Santos-Silva, António
Alvarez, Ezequiel
Correia, Ilídio Joaquim Sobreira
Verde, Ignacio
author_role author
author2 Santos-Silva, António
Alvarez, Ezequiel
Correia, Ilídio Joaquim Sobreira
Verde, Ignacio
author2_role author
author
author
author
dc.contributor.none.fl_str_mv uBibliorum
dc.contributor.author.fl_str_mv Cairrão, Elisa
Santos-Silva, António
Alvarez, Ezequiel
Correia, Ilídio Joaquim Sobreira
Verde, Ignacio
dc.subject.por.fl_str_mv Umbilical artery
Smooth muscle cells
Patch-clamp
Calcium channels
topic Umbilical artery
Smooth muscle cells
Patch-clamp
Calcium channels
description The human umbilical cord is a biological sample that can be easily obtained just after birth. A methodology was developed to perform cultures of human umbilical artery smooth muscle cells (HUASMC) expressing contractile proteins and functional ionic channels. To avoid fibroblast and endothelial cell contamination, we mechanically separated the tunica media, which only contains HUASMC and matrix proteins. To isolate the cells, collagenase V and elastase were used as hydrolyzing enzymes. The isolated cells were plated in collagen-coated dishes to obtain cultures of HUASMC. The cells obtained after different passages (1 to 6) exhibit the characteristic vascular smooth cell morphology and express smooth muscle alpha-2 actin, myosin heavy chain SM1, and alpha subunits of L- and T-type calcium channels (Cav 1.2, Cav 1.2, and Cav 3.2). Electrophysiology recordings for L- and T-type calcium channels were made, indicating that these channels are functional in the cultured cells. In conclusion, the procedure developed allows obtaining cultures of HUASMC expressing contractile proteins and also functional ionic channels. These cells could be used to study cellular and molecular aspects about the regulation of the vascular function.
publishDate 2009
dc.date.none.fl_str_mv 2009-01-01
2009-01-01T00:00:00Z
2018-03-15T16:00:29Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.6/4622
url http://hdl.handle.net/10400.6/4622
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Cairrão, E., Santos-Silva, A.J., Alvarez, E., Correia, I.J. e Verde, I. (2009) “Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels”, In Vitro Cellular & Developmental Biology – Animal, Vol. 45 (3), pp. 175-184
10.1007/s11626-008-9161-6
dc.rights.driver.fl_str_mv metadata only access
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dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
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