Mouse corneal lymphangiogenesis model

• Published in: Nature protocols Vol. 6; no. 6; pp. 817 - 826
• Main Authors: Cao, Renhai, Lim, Sharon, Ji, Hong, Zhang, Yin, Yang, Yunlong, Honek, Jennifer, Hedlund, Eva-Maria, Cao, Yihai
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-06-2011; Nature Publishing Group
• Subjects: 631/136/2060/290; 631/1647/664/1257; Analytical Chemistry; Angiogenesis; Animals; Biological Techniques; Biomedical and Life Sciences; Cancer; Computational Biology/Bioinformatics; Cornea; Cornea - pathology; Cornea - physiology; Cytokines; Hyaluronic acid; Immunohistochemistry; Inflammation; Life Sciences; Lymphangiogenesis; Lymphatic Vessels - pathology; Lymphatic Vessels - physiology; MEDICIN; Medicin och hälsovetenskap; MEDICINE; Mice; Microarrays; Models, Animal; Organic Chemistry; Physiological aspects; Polymers; protocol; Sutures; Vascular endothelial growth factor; Sweden
• ISSN: 1754-2189; 1750-2799; 1750-2799
• DOI: 10.1038/nprot.2011.359

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.