Human breast microvascular endothelial cells retain phenotypic traits in long-term finite life span culture.

2.50
Hdl Handle:
http://hdl.handle.net/2336/18694
Title:
Human breast microvascular endothelial cells retain phenotypic traits in long-term finite life span culture.
Authors:
Sigurdsson, Valgardur; Fridriksdottir, Agla J R; Kjartansson, Jens; Jonasson, Jon G; Steinarsdottir, Margret; Petersen, Ole William; Ogmundsdottir, Helga M; Gudjonsson, Thorarinn
Citation:
In Vitro Cell Dev Biol Anim., 42(10):332-40
Issue Date:
1-Dec-2006
Abstract:
Attempts to study endothelial-epithelial interactions in the human breast have been hampered by lack of protocols for long-term cultivation of breast endothelial cells (BRENCs). The aim of this study was to establish long-term cultures of BRENCs and to compare their phenotypic traits with the tissue of origin. Microvasculature was localized in situ by immunohistochemistry in breast samples. From this tissue, collagen-rich stroma and adipose tissue were dissected mechanically and further disaggregated to release microvessel organoids. BRENCs were cultured from these organoids in endothelial specific medium and characterized by staining for endothelial markers. Microvessels were a prominent feature of intralobular tissue as evidenced by immunostaining against endothelial specific markers such as CD31, VE-cadherin, and von Willebrand factor (VWF). Double staining against VE-cadherin and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) showed that blood and lymphatic vessels could be distinguished. An antibody against CD31 was used to refine protocols for isolation of microvasculature from reduction mammoplasties. BRENCs retained critical traits even at high passage, including uptake of low-density lipoprotein, and had E-selectin induced upon treatment with tumor necrosis factor-alpha. The first signs of senescence in passage 14 were accompanied by gain of trisomy 11. At passage 18 cells showed chromosomal aberrations and growth arrest as revealed by beta-galactosidase staining. We demonstrate here that breast microvasculature may serve as a large-scale source for expansion of BRENCs with molecular and functional traits preserved. These cells will form the basis for studies on the role of endothelial cells in breast morphogenesis.
Description:
To access publisher full text version of this article. Please click on the hyperlink in Additional Links field
Additional Links:
http://www.springerlink.com/content/v5v3019337562817

Full metadata record

DC FieldValue Language
dc.contributor.authorSigurdsson, Valgardur-
dc.contributor.authorFridriksdottir, Agla J R-
dc.contributor.authorKjartansson, Jens-
dc.contributor.authorJonasson, Jon G-
dc.contributor.authorSteinarsdottir, Margret-
dc.contributor.authorPetersen, Ole William-
dc.contributor.authorOgmundsdottir, Helga M-
dc.contributor.authorGudjonsson, Thorarinn-
dc.date.accessioned2008-02-20T09:56:47Z-
dc.date.available2008-02-20T09:56:47Z-
dc.date.issued2006-12-01-
dc.date.submitted2008-02-20-
dc.identifier.citationIn Vitro Cell Dev Biol Anim., 42(10):332-40en
dc.identifier.issn1071-2690-
dc.identifier.pmid17316068-
dc.identifier.doi10.1290/0602017.1-
dc.identifier.urihttp://hdl.handle.net/2336/18694-
dc.descriptionTo access publisher full text version of this article. Please click on the hyperlink in Additional Links fielden
dc.description.abstractAttempts to study endothelial-epithelial interactions in the human breast have been hampered by lack of protocols for long-term cultivation of breast endothelial cells (BRENCs). The aim of this study was to establish long-term cultures of BRENCs and to compare their phenotypic traits with the tissue of origin. Microvasculature was localized in situ by immunohistochemistry in breast samples. From this tissue, collagen-rich stroma and adipose tissue were dissected mechanically and further disaggregated to release microvessel organoids. BRENCs were cultured from these organoids in endothelial specific medium and characterized by staining for endothelial markers. Microvessels were a prominent feature of intralobular tissue as evidenced by immunostaining against endothelial specific markers such as CD31, VE-cadherin, and von Willebrand factor (VWF). Double staining against VE-cadherin and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) showed that blood and lymphatic vessels could be distinguished. An antibody against CD31 was used to refine protocols for isolation of microvasculature from reduction mammoplasties. BRENCs retained critical traits even at high passage, including uptake of low-density lipoprotein, and had E-selectin induced upon treatment with tumor necrosis factor-alpha. The first signs of senescence in passage 14 were accompanied by gain of trisomy 11. At passage 18 cells showed chromosomal aberrations and growth arrest as revealed by beta-galactosidase staining. We demonstrate here that breast microvasculature may serve as a large-scale source for expansion of BRENCs with molecular and functional traits preserved. These cells will form the basis for studies on the role of endothelial cells in breast morphogenesis.en
dc.language.isoenen
dc.publisherSociety for In Vitro Biologyen
dc.relation.urlhttp://www.springerlink.com/content/v5v3019337562817en
dc.subject.meshBreasten
dc.subject.meshCell Agingen
dc.subject.meshCell Separationen
dc.subject.meshCells, Cultureden
dc.subject.meshChromosomal Instabilityen
dc.subject.meshEndothelial Cellsen
dc.subject.meshFemaleen
dc.subject.meshHumansen
dc.subject.meshKaryotypingen
dc.subject.meshPhenotypeen
dc.subject.meshTime Factorsen
dc.titleHuman breast microvascular endothelial cells retain phenotypic traits in long-term finite life span culture.en
dc.typeArticleen
dc.identifier.eissn1543-706X-
dc.contributor.departmentFaculty of Medicine, University of Iceland, Reykjavik, Iceland.en
dc.identifier.journalIn vitro cellular & developmental biology. Animalen

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