Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Outros Autores: | , , , |
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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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 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
metadata only access |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Springer |
publisher.none.fl_str_mv |
Springer |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799136354396798976 |