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An improved method for isolation of microvascular endothelial cells from normal and inflamed human lung

Lou, Jin ; Mili, Nabil ; Decrind, Christine ; Donati, Yves ; Kossodo, Sylvie ; Spiliopoulos, Anastase ; Ricou, Bara ; Suter, Peter ; Morel, Denis ; Morel, Philippe ; Grau, Georges

In: In Vitro Cellular & Developmental Biology - Animal, 1998, vol. 34, no. 7, p. 529-536

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    Titre
    • An improved method for isolation of microvascular endothelial cells from normal and inflamed human lung
    Auteur
    • Lou, Jin. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Mili, Nabil. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Decrind, Christine. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Donati, Yves. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Kossodo, Sylvie. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Spiliopoulos, Anastase. Division of Surgical Intensive Care, Clinics of Cardiovascular and Thoracic Surgery, University Hospital and University Medical Center, CH-1211, Geneva 4, Switzerland
    • Ricou, Bara. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Suter, Peter. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Morel, Denis. Division of Investigative Surgery, Department of Surgery, University Hospital, University of Geneva, 24, rue Micheli-du-crest 1211, Geneva 14, Switzerland
    • Morel, Philippe. Division of Investigative Surgery, Department of Surgery, University Hospital and University Medical Center, CH-1211, Geneva 4, Switzerland
    • Grau, Georges. Division of Surgical Intensive Care, Clinics of Cardiovascular and Thoracic Surgery, University Hospital and University Medical Center, CH-1211, Geneva 4, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • In Vitro Cellular & Developmental Biology - Animal, 1998, vol. 34, no. 7, p. 529-536. Springer-Verlag
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:314647
    Summary
    Summary: Microvascular endothelial cells (MVEC), which differ from large vessel endothelial cells, have been isolated successfully from lungs of various species, including man. However, contamination by nonendothelial cells remains a major problem in spite of several technical improvements. In view of the organ specificity of MVEC, endothelial cells should be derived from the tissue involved in the diseases one wishes to study. Therefore, to investigate some of the immunopathological mechanisms leading to acute respiratory distress syndrome (ARDS), we have attempted to isolate lung MVEC from patients undergoing thoracic surgery for lung carcinoma and patients dying of ARDS. The method described here includes four main steps: (1) full digestion of pulmonary tissue with trypsin and collagenase, (2) aggregation of MVEC induced by human plasma, (3) Percoll density centrifugation, and (4) selection and transfer of MVEC after local digestion with trypsin/EDTA under light microscopy. Normal and ARDS-derived lung MVEC purified by this technique presented contact inhibition (i.e., grew in monolayer), and expressed classical endothelial markers, including von Willebrand factor (vWF), platelet endothelial cell adhesion molecule 1(PECAM-1, CD31), and transcripts for the angiogensin converting enzyme (ACE). The cells also formed capillarylike structures, took up high levels of acetylated low-density lipoprotein (Ac-LDL), and exhibited ELAM-1 inducibility in response to TNF. Contaminant cells, such as fibroblasts, smooth muscle cells, or pericytes, were easily recognized on the basis of morphology and were eliminated by selection of plasma-aggregated cells under light microscopy. The technique presented here allows one to study the specific involvement and contribution of pulmonary endothelium in various lung diseases