title: Cell-based approach for 3D reconstruction of lymphatic capillaries in vitro reveals distinct functions of HGF and VEGF-C in lymphangiogenesis.
abstract: Regeneration of lymphatic vessels is important for treatment of various disorders of lymphatic system and for restoration of lymphatic function after surgery. We have developed a method for generating a human 3D lymphatic vascular construct. In this system, human lymphatic endothelial cells, co-cultured with fibroblasts, spontaneously organized into a stable 3D lymphatic capillary network without the use of any exogenous factors. In vitro-generated lymphatic capillaries exhibited the major molecular and ultra-structural features of native, human lymphatic microvasculature: branches in the three dimensions, wide lumen, blind ends, overlapping borders, adherens and tight junctions, anchoring filaments, lack of mural cells, and poorly developed basement membrane. Furthermore, we show that fibroblast-derived VEGF-C and HGF cooperate in the formation of lymphatic vasculature by activating ERK1/2 signaling, and demonstrate distinct functions of HGF/c-Met and VEGF-C/VEGFR-3 in lymphangiogenesis. This lymphatic vascular construct is expected to facilitate studies of lymphangiogenesis in vitro and it holds promise as a strategy for regeneration of lymphatic vessels and treatment of lymphatic disorders in various conditions.
authors.affiliation: Centre LOEX de l'Université Laval, Regenerative Medicine section of the FRQS Research, Center of the CHU de Québec, Quebec G1J1Z4, QC, Canada.
authors.forename: Laure
authors.initials: L
authors.lastname: Gibot
authors.affiliation: Centre LOEX de l'Université Laval, Regenerative Medicine section of the FRQS Research, Center of the CHU de Québec, Quebec G1J1Z4, QC, Canada.
authors.forename: Todd
authors.initials: T
authors.lastname: Galbraith
authors.affiliation: Department of Oncological Sciences and Tisch Cancer Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Bryan
authors.initials: B
authors.lastname: Kloos
authors.affiliation: Department of Oncological Sciences and Tisch Cancer Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Suvendu
authors.initials: S
authors.lastname: Das
authors.affiliation: Centre LOEX de l'Université Laval, Regenerative Medicine section of the FRQS Research, Center of the CHU de Québec, Quebec G1J1Z4, QC, Canada.
authors.forename: Dan A
authors.initials: DA
authors.lastname: Lacroix
authors.affiliation: Centre LOEX de l'Université Laval, Regenerative Medicine section of the FRQS Research, Center of the CHU de Québec, Quebec G1J1Z4, QC, Canada.
authors.forename: François A
authors.initials: FA
authors.lastname: Auger
authors.affiliation: Department of Oncological Sciences and Tisch Cancer Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: mihaela.skobe@mssm.edu.
authors.forename: Mihaela
authors.initials: M
authors.lastname: Skobe
mesh_terms: D017228:Hepatocyte Growth Factor / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D042601:Lymphatic Vessels / Q000033:anatomy & histology*; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism*
chemical_list: D042582:Vascular Endothelial Growth Factor C; D017228:Hepatocyte Growth Factor
keywords: 3D; HGF; Lymphangiogenesis; Lymphatic capillary; Self-assembly; VEGF-C; VEGFR-3; c-Met