Isolation and Characterization of Human Lung Lymphatic Endothelial Cells

人肺淋巴管内皮细胞的分离和鉴定

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作者:Bruno Lorusso, Angela Falco, Denise Madeddu, Caterina Frati, Stefano Cavalli, Gallia Graiani, Andrea Gervasi, Laura Rinaldi, Costanza Lagrasta, Davide Maselli, Letizia Gnetti, Enrico M Silini, Eugenio Quaini, Luca Ampollini, Paolo Carbognani, Federico Quaini

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 in vitro 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.

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