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

Article Article scientifique Article An improved method for isolation of microvascular endothelial cells from normal and inflamed human lung eng 1998 author Lou Jinning 26933 author Mili Nabil 63021 author Decrind Christine author Donati Yves Richard 24165 author Kossodo Sylvie 21916 author Spiliopoulos Anastase 49910 author Ricou Bara 127716 author Suter Peter 25405 author Morel Denis 22755 author Morel Philippe 22337 author Grau Georges 22432 529-536 10.1007/s11626-998-0112-z 9719412 1071-2690 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. Antigens, CD31/metabolism Base Sequence Cytological Techniques/standards DNA Primers E-Selectin/metabolism Endothelium, Vascular/cytology/metabolism/pathology Humans Lipoproteins, LDL/metabolism Lung/blood supply Peptidyl-Dipeptidase A/genetics RNA, Messenger/genetics/metabolism Respiratory Distress Syndrome, Adult/pathology Tumor Necrosis Factor-alpha/physiology Von Willebrand Factor/metabolism Dewey 616.07 Dewey 617 137 Département de pathologie et immunologie 21F 57 Département de chirurgie 222 63 Département d'anesthésiologie, pharmacologie, soins intensifs et urgences 229 false 79 Chirurgie viscérale 104 1329 Chirurgie viscérale (HUG) 222 Devenir des patients ayant séjourné aux soins intensifs de chirurgie 283 In vitro cellular & developmental biology. Animal 34 7 d863caf3-63cb-4071-8fbb-0563fda705b1 Article (Published version) Public art%253A10.1007%252Fs11626-998-0112-z.pdf 2470134 application/pdf openAccess Faculté de médecine / Section de médecine fondamentale / Département de pathologie et immunologie Faculté de médecine / Section de médecine clinique / Département de chirurgie Faculté de médecine / Section de médecine clinique / Département d'anesthésiologie, pharmacologie, soins intensifs et urgences

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An improved method for isolation of microvascular endothelial cells from normal and inflamed human lung	In vitro cellular & developmental biology. Animal			1998	830	604

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Kossodo, Sylvie