Isolation and Characterization of Human Lung Lymphatic Endothelial Cells
Identifieur interne : 001C28 ( Main/Exploration ); précédent : 001C27; suivant : 001C29Isolation and Characterization of Human Lung Lymphatic Endothelial Cells
Auteurs : Bruno Lorusso [Italie] ; Angela Falco [Italie] ; Denise Madeddu [Italie] ; Caterina Frati [Italie] ; Stefano Cavalli [Italie] ; Gallia Graiani [Italie] ; Andrea Gervasi [Italie] ; Laura Rinaldi [Italie] ; Costanza Lagrasta [Italie] ; Davide Maselli [Italie] ; Letizia Gnetti [Italie] ; Enrico M. Silini [Italie] ; Eugenio Quaini [Italie] ; Luca Ampollini [Italie] ; Paolo Carbognani [Italie] ; Federico Quaini [Italie]Source :
- BioMed Research International [ 2314-6133 ] ; 2015.
Abstract
Characterization of lymphatic endothelial cells from the respiratory system may be crucial to investigate the role of the lymphatic system in the normal and diseased lung. We describe a simple and inexpensive method to harvest, isolate, and expand lymphatic endothelial cells from the human lung (HL-LECs). Fifty-five samples of healthy lung selected from patients undergoing lobectomy were studied. A two-step purification tool, based on paramagnetic sorting with monoclonal antibodies to CD31 and Podoplanin, was employed to select a pure population of HL-LECs. The purity of HL-LECs was assessed by morphologic criteria, immunocytochemistry, flow cytometry, and functional assays. Interestingly, these cells retain
Url:
DOI: 10.1155/2015/747864
PubMed: 26137493
PubMed Central: 4475539
Affiliations:
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wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Lagrasta, Costanza" sort="Lagrasta, Costanza" uniqKey="Lagrasta C" first="Costanza" last="Lagrasta">Costanza Lagrasta</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Maselli, Davide" sort="Maselli, Davide" uniqKey="Maselli D" first="Davide" last="Maselli">Davide Maselli</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Gnetti, Letizia" sort="Gnetti, Letizia" uniqKey="Gnetti L" first="Letizia" last="Gnetti">Letizia Gnetti</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of 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uniqKey="Lorusso B" first="Bruno" last="Lorusso">Bruno Lorusso</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Falco, Angela" sort="Falco, Angela" uniqKey="Falco A" first="Angela" last="Falco">Angela Falco</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Madeddu, Denise" sort="Madeddu, Denise" uniqKey="Madeddu D" first="Denise" last="Madeddu">Denise Madeddu</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Frati, Caterina" sort="Frati, Caterina" uniqKey="Frati C" first="Caterina" last="Frati">Caterina Frati</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Cavalli, Stefano" sort="Cavalli, Stefano" uniqKey="Cavalli S" first="Stefano" last="Cavalli">Stefano Cavalli</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Graiani, Gallia" sort="Graiani, Gallia" uniqKey="Graiani G" first="Gallia" last="Graiani">Gallia Graiani</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Gervasi, Andrea" sort="Gervasi, Andrea" uniqKey="Gervasi A" first="Andrea" last="Gervasi">Andrea Gervasi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Rinaldi, Laura" sort="Rinaldi, Laura" uniqKey="Rinaldi L" first="Laura" last="Rinaldi">Laura Rinaldi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Lagrasta, Costanza" sort="Lagrasta, Costanza" uniqKey="Lagrasta C" first="Costanza" last="Lagrasta">Costanza Lagrasta</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Maselli, Davide" sort="Maselli, Davide" uniqKey="Maselli D" first="Davide" last="Maselli">Davide Maselli</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Gnetti, Letizia" sort="Gnetti, Letizia" uniqKey="Gnetti L" first="Letizia" last="Gnetti">Letizia Gnetti</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Silini, Enrico M" sort="Silini, Enrico M" uniqKey="Silini E" first="Enrico M." last="Silini">Enrico M. Silini</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Biomedical, Biotechnological, and Translational Sciences (S.Bi.Bi.T.), University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Quaini, Eugenio" sort="Quaini, Eugenio" uniqKey="Quaini E" first="Eugenio" last="Quaini">Eugenio Quaini</name><affiliation wicri:level="1"><nlm:aff id="I3">Cardiovascular Department, Humanitas Clinical and Research Centre, 20089 Milan, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Cardiovascular Department, Humanitas Clinical and Research Centre, 20089 Milan</wicri:regionArea><placeName><settlement type="city">Milan</settlement><region nuts="2">Lombardie</region></placeName></affiliation></author><author><name sortKey="Ampollini, Luca" sort="Ampollini, Luca" uniqKey="Ampollini L" first="Luca" last="Ampollini">Luca Ampollini</name><affiliation wicri:level="1"><nlm:aff id="I4">Department of Surgical Sciences, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Surgical Sciences, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Carbognani, Paolo" sort="Carbognani, Paolo" uniqKey="Carbognani P" first="Paolo" last="Carbognani">Paolo Carbognani</name><affiliation wicri:level="1"><nlm:aff id="I4">Department of Surgical Sciences, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Surgical Sciences, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author><author><name sortKey="Quaini, Federico" sort="Quaini, Federico" uniqKey="Quaini F" first="Federico" last="Quaini">Federico Quaini</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Clinical and Experimental Medicine, University Hospital of Parma, 43126 Parma</wicri:regionArea><wicri:noRegion>43126 Parma</wicri:noRegion></affiliation></author></analytic><series><title level="j">BioMed Research International</title><idno type="ISSN">2314-6133</idno><idno type="eISSN">2314-6141</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Characterization of lymphatic endothelial cells from the respiratory system may be crucial to investigate the role of the lymphatic system in the normal and diseased lung. We describe a simple and inexpensive method to harvest, isolate, and expand lymphatic endothelial cells from the human lung (HL-LECs). Fifty-five samples of healthy lung selected from patients undergoing lobectomy were studied. A two-step purification tool, based on paramagnetic sorting with monoclonal antibodies to CD31 and Podoplanin, was employed to select a pure population of HL-LECs. The purity of HL-LECs was assessed by morphologic criteria, immunocytochemistry, flow cytometry, and functional assays. Interestingly, these cells retain <italic>in vitro</italic> several receptor tyrosine kinases (RTKs) implicated in cell survival and proliferation. HL-LECs represent a clinically relevant cellular substrate to study lymphatic biology, lymphoangiogenesis, interaction with microbial agents, wound healing, and anticancer therapy.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Oliver, G" uniqKey="Oliver G">G. 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Aird</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Italie</li></country><region><li>Lombardie</li></region><settlement><li>Milan</li></settlement></list><tree><country name="Italie"><noRegion><name sortKey="Lorusso, Bruno" sort="Lorusso, Bruno" uniqKey="Lorusso B" first="Bruno" last="Lorusso">Bruno Lorusso</name></noRegion><name sortKey="Ampollini, Luca" sort="Ampollini, Luca" uniqKey="Ampollini L" first="Luca" last="Ampollini">Luca Ampollini</name><name sortKey="Carbognani, Paolo" sort="Carbognani, Paolo" uniqKey="Carbognani P" first="Paolo" last="Carbognani">Paolo Carbognani</name><name sortKey="Cavalli, Stefano" sort="Cavalli, Stefano" uniqKey="Cavalli S" first="Stefano" last="Cavalli">Stefano Cavalli</name><name sortKey="Falco, Angela" sort="Falco, Angela" uniqKey="Falco A" first="Angela" last="Falco">Angela Falco</name><name sortKey="Frati, Caterina" sort="Frati, Caterina" uniqKey="Frati C" first="Caterina" last="Frati">Caterina Frati</name><name sortKey="Gervasi, Andrea" sort="Gervasi, Andrea" uniqKey="Gervasi A" first="Andrea" last="Gervasi">Andrea Gervasi</name><name sortKey="Gnetti, Letizia" sort="Gnetti, Letizia" uniqKey="Gnetti L" first="Letizia" last="Gnetti">Letizia Gnetti</name><name sortKey="Graiani, Gallia" sort="Graiani, Gallia" uniqKey="Graiani G" first="Gallia" last="Graiani">Gallia Graiani</name><name sortKey="Lagrasta, Costanza" sort="Lagrasta, Costanza" uniqKey="Lagrasta C" first="Costanza" last="Lagrasta">Costanza Lagrasta</name><name sortKey="Madeddu, Denise" sort="Madeddu, Denise" uniqKey="Madeddu D" first="Denise" last="Madeddu">Denise Madeddu</name><name sortKey="Maselli, Davide" sort="Maselli, Davide" uniqKey="Maselli D" first="Davide" last="Maselli">Davide Maselli</name><name sortKey="Quaini, Eugenio" sort="Quaini, Eugenio" uniqKey="Quaini E" first="Eugenio" last="Quaini">Eugenio Quaini</name><name sortKey="Quaini, Federico" sort="Quaini, Federico" uniqKey="Quaini F" first="Federico" last="Quaini">Federico Quaini</name><name sortKey="Rinaldi, Laura" sort="Rinaldi, Laura" uniqKey="Rinaldi L" first="Laura" last="Rinaldi">Laura Rinaldi</name><name sortKey="Silini, Enrico M" sort="Silini, Enrico M" uniqKey="Silini E" first="Enrico M." last="Silini">Enrico M. 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