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

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Published in In vitro cellular & developmental biology. Animal. 1998, vol. 34, no. 7, p. 529-36
Collection Open Access - Licence nationale Springer
Abstract 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 angiotensin 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.
Keywords Antigens, CD31/metabolismBase SequenceCytological Techniques/standardsDNA PrimersE-Selectin/metabolismEndothelium, Vascular/cytology/metabolism/pathologyHumansLipoproteins, LDL/metabolismLung/blood supplyPeptidyl-Dipeptidase A/geneticsRNA, Messenger/genetics/metabolismRespiratory Distress Syndrome, Adult/pathologyTumor Necrosis Factor-alpha/physiologyVon Willebrand Factor/metabolism
PMID: 9719412
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Research groups Chirurgie viscérale (104)
Chirurgie viscérale (HUG)
Devenir des patients ayant séjourné aux soins intensifs de chirurgie (283)
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LOU, Jinning et al. An improved method for isolation of microvascular endothelial cells from normal and inflamed human lung. In: In vitro cellular & developmental biology. Animal, 1998, vol. 34, n° 7, p. 529-36. doi: 10.1007/s11626-998-0112-z https://archive-ouverte.unige.ch/unige:74239

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Deposited on : 2015-07-22

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