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An affordable method to obtain cultured endothelial cells from peripheral blood

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An affordable method to obtain cultured endothelial cells from peripheral blood

dc.contributor.author	Bueno Betí, Carlos
dc.contributor.author	Novella del Campo, Susana
dc.contributor.author	Lázaro-Franco, Macarena
dc.contributor.author	Heras, Magda
dc.contributor.author	Pérez Cremades, Daniel
dc.contributor.author	Sanchís, Juan
dc.contributor.author	Hermenegildo, Carlos
dc.date.accessioned	2015-05-07T11:49:48Z
dc.date.available	2015-05-07T11:49:48Z
dc.date.issued	2013
dc.identifier.uri	http://hdl.handle.net/10550/43595
dc.description.abstract	The culture of endothelial progenitor cells (EPC) provides an excellent tool to research on EPC biology and vascular regeneration and vasculogenesis. The use of different protocols to obtain EPC cultures makes it difficult to obtain comparable results in different groups. This work offers a systematic comparison of the main variables of most commonly used protocols for EPC isolation, culture and functional evaluation. Peripheral blood samples from healthy individuals were recovered and mononuclear cells were cultured. Different recovery and culture conditions were tested: blood volume, blood anticoagulant, coating matrix and percentage of foetal bovine serum (FBS) in culture media. The success of culture procedure, first colonies of endothelial cells appearance time, correlation with number of circulating EPC (cEPC) and functional comparison with human umbilical vein endothelial cells (HUVEC) were studied. The use of heparin, a minimum blood volume of 30 ml, fibronectin as a coating matrix and endothelial growing media-2 supplemented with 20% FBS increased the success of obtaining EPC cultures up to 80% of the processed samples while reducing EPC colony appearance mean time to a minimum of 13 days. Blood samples exhibiting higher cEPC numbers resulted in reduced EPC colony appearance mean time. Cells isolated by using this combination were endothelial cell-like EPCs morphological and phenotypically. Functionally, cultured EPC showed decreased growing and vasculogenic capacity when compared to HUVEC. Thus, above-mentioned conditions allow the isolation and culture of EPC with smaller blood volumes and shorter times than currently used protocols.
dc.language.iso	eng
dc.relation.ispartof	Journal Of Cellular And Molecular Medicine, 2013, vol. 17, num. 11, p. 1475-1483
dc.rights.uri	info:eu-repo/semantics/openAccess
dc.source	Bueno Betí, Carlos Novella, Susaqna Lázaro Franco, Macarena Pérez Cremades, Daniel Heras, Magda Sanchís, Juan Hermenegildo, Carlos 2013 An affordable method to obtain cultured endothelial cells from peripheral blood. Journal Of Cellular And Molecular Medicine 17 11 1475 1483
dc.subject	Fisiologia cel·lular
dc.subject	Sang
dc.title	An affordable method to obtain cultured endothelial cells from peripheral blood
dc.type	info:eu-repo/semantics/article
dc.date.updated	2015-05-07T11:49:48Z
dc.identifier.doi	http://dx.doi.org/10.1111/jcmm.12133
dc.identifier.idgrec	093136