title: Mouse corneal lymphangiogenesis model.
abstract: This protocol describes a powerful in vivo method to quantitatively study the formation of new lymphatic vessels in the avascular cornea without interference of pre-existing lymphatics. Implantation of 100 ng of lymphangiogenic factors such as vascular endothelial growth factor (VEGF)-A, VEGF-C or fibroblast growth factor-2, together with slow-release polymers, into a surgically created micropocket in the mouse cornea elicits a robust lymphangiogenic response. Newly formed lymphatic vessels are detected by immunohistochemical staining of the flattened corneal tissue with lymphatic endothelial-specific markers such as lymphatic vessel endothelial hyaluronan receptor-1; less-specific markers such as vascular endothelial growth factor receptor 3 may also be used. Lymphatic vessel growth in relation to hemangiogenesis can be readily detected starting at day 5 or 6 after pellet implantation and persists for ∼14 d. This protocol offers a unique opportunity to study the mechanisms underlying lymphatic vessel formation, remodeling and function.
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Renhai
authors.initials: R
authors.lastname: Cao
authors.forename: Sharon
authors.initials: S
authors.lastname: Lim
authors.forename: Hong
authors.initials: H
authors.lastname: Ji
authors.forename: Yin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Yang
authors.forename: Jennifer
authors.initials: J
authors.lastname: Honek
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Hedlund
authors.forename: Yihai
authors.initials: Y
authors.lastname: Cao
mesh_terms: D000818:Animals; D003315:Cornea / Q000473:pathology / Q000502:physiology*; D007150:Immunohistochemistry; D042583:Lymphangiogenesis; D042601:Lymphatic Vessels / Q000473:pathology / Q000502:physiology*; D051379:Mice; D023421:Models, Animal