Abstract: Objective: To assist further investigation of the function of lymphangiogenesis during cancer metastasis. Methods: Hematoxylin-eosine staining and immunohistochemistry were used to detect histopathological features of lymphangioma. Human lymphatic endothelial cells were isolated by collagenase digestion, and were cultured in M199 medium added with vascular growth factor and heparin. Morphology and ultrastructure of cells were observed under contras phase light microscope and transmission electron microscope. MTT was applied to survey proliferation of lymphatic endothelial cells. Results: Hematoxylin-eosine staining sections revealed that numerous dilated vessels were generally presence in glossal lymphanigoma specimen. FLT-4 and LYVE-1 were positive in lymphatic endothelial cells. LECs showed a typical cobblestone appearance observed under contrast phase light microphage. The typical granule, weibel-palade body, that was unique to the endothelial cells, is exhibited in the ultrastructure of lymphatic endothelial cells. Gap intercellular junctions in adjacent lymphatic endothelial cells were overviewed in the ultrathin sections of lymphatic endothelial cells. Lymphatic endothelial cells formed tube-like structures and communicated with each other in the collagenⅠgel to form a complex vascular plexus similar to the vessel system in vivo. Conclusion: We have successfully isolated and expanded LECs in vitro, and provided a favorable model for researching the phenotypic and functional properties of LECs.

Key words: Lymphangioma , Lymphatic endothelial cell, Cell culture