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An affordable method to obtain cultured endothelial cells from peripheral blood.

Bueno-Betí, C; Novella, S; Lázaro-Franco, M; Pérez-Cremades, D; Heras, M; Sanchís, J; Hermenegildo, C (2013) An affordable method to obtain cultured endothelial cells from peripheral blood. J Cell Mol Med, 17 (11). pp. 1475-1483. ISSN 1582-4934 https://doi.org/10.1111/jcmm.12133
SGUL Authors: Bueno Beti, Carlos

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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.

Item Type: Article
Additional Information: © 2013 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine. This is an open access article under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: cell culture, endothelial progenitor cells, vasculogenesis, Blood Cells, Cell Adhesion, Cell Culture Techniques, Cell Proliferation, Cell Separation, Cells, Cultured, Human Umbilical Vein Endothelial Cells, Humans, Neovascularization, Physiologic, Reproducibility of Results, Stem Cells, Blood Cells, Cells, Cultured, Stem Cells, Humans, Cell Culture Techniques, Cell Separation, Reproducibility of Results, Cell Adhesion, Cell Proliferation, Neovascularization, Physiologic, Human Umbilical Vein Endothelial Cells, endothelial progenitor cells, cell culture, vasculogenesis, 0304 Medicinal and Biomolecular Chemistry, 0601 Biochemistry and Cell Biology, 1103 Clinical Sciences, Biochemistry & Molecular Biology
SGUL Research Institute / Research Centre: Academic Structure > Molecular and Clinical Sciences Research Institute (MCS)
Journal or Publication Title: J Cell Mol Med
ISSN: 1582-4934
Language: eng
Dates:
DateEvent
23 December 2013Published
1 October 2013Published Online
14 August 2013Accepted
Publisher License: Creative Commons: Attribution 3.0
PubMed ID: 24118735
Web of Science ID: WOS:000328693900013
Go to PubMed abstract
URI: https://openaccess.sgul.ac.uk/id/eprint/111981
Publisher's version: https://doi.org/10.1111/jcmm.12133

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