title: A Methodology for Concomitant Isolation of Intimal and Adventitial Endothelial Cells from the Human Thoracic Aorta.
abstract: BACKGROUND
  Aortic diseases are diverse and involve a multiplicity of biological systems in the vascular wall. Aortic dissection, which is usually preceded by aortic aneurysm, is a leading cause of morbidity and mortality in modern societies. Although the endothelium is now known to play an important role in vascular diseases, its contribution to aneurysmal aortic lesions remains largely unknown. The aim of this study was to define a reliable methodology for the isolation of aortic intimal and adventitial endothelial cells in order to throw light on issues relevant to endothelial cell biology in aneurysmal diseases.
  
  
  RESULTS
  We set up protocols to isolate endothelial cells from both the intima and the adventitia of human aneurysmal aortic vessel segments. Throughout the procedure, analysis of cell morphology and endothelial markers allowed us to select an endothelial fraction which after two rounds of expansion yielded a population of >90% pure endothelial cells. These cells have the features and functionalities of freshly isolated cells and can be used for biochemical studies. The technique was successfully used for aortic vessel segments of 20 patients and 3 healthy donors.
  
  
  CONCLUSIONS
  This simple and highly reproducible method allows the simultaneous preparation of reasonably pure primary cultures of intimal and adventitial human endothelial cells, thus providing a reliable source for investigating their biology and involvement in both thoracic aneurysms and other aortic diseases.
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM, U1045, Bordeaux, France.|Assistance Publique-Hôpitaux de Paris, Hôpital Bichat, Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Leclercq
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM, U1045, Bordeaux, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Veillat
authors.affiliation: Université de Bordeaux, Bordeaux, France.|SFR TransBioMed, Bordeaux, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Loriot
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM, U1045, Bordeaux, France.
authors.forename: Pirjo
authors.initials: P
authors.lastname: Spuul
authors.affiliation: Service de chirurgie cardiaque et vasculaire, Hôpital Haut-L'Evêque, Pessac, France.
authors.forename: Francesco
authors.initials: F
authors.lastname: Madonna
authors.affiliation: Service de chirurgie cardiaque et vasculaire, Hôpital Haut-L'Evêque, Pessac, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Roques
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM, U1045, Bordeaux, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Génot
mesh_terms: D001013:Aorta, Thoracic / Q000378:metabolism / Q000473:pathology*; D001014:Aortic Aneurysm / Q000473:pathology / Q000601:surgery; D015415:Biomarkers; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology*; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D016130:Immunophenotyping; D010641:Phenotype; D061251:Primary Cell Culture; D015203:Reproducibility of Results; D017539:Tunica Intima / Q000378:metabolism / Q000473:pathology*
chemical_list: D015415:Biomarkers