A simple method for culturing mouse vascular endothelium

• Published in: Journal of immunological methods 2001-08, Vol.254 (1), p.31-45
• Main Authors: Kreisel, Daniel, Krupnick, Alexander S, Szeto, Wilson Y, Popma, Sicco H, Sankaran, David, Krasinskas, Alyssa M, Amin, Kunjlata M, Rosengard, Bruce R
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD - biosynthesis; Aorta, Thoracic - cytology; Biological and medical sciences; Biomarkers; Blotting, Western; CD4-Positive T-Lymphocytes - metabolism; CD8-Positive T-Lymphocytes - metabolism; Cell Culture Techniques - methods; Cells, Cultured; Culture methods; Diverse techniques; Endothelial cells; Endothelium, Vascular - cytology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; g-Interferon; Lipoproteins, LDL - metabolism; Male; Mice; Mice, Inbred C57BL; Microbiology; Microscopy, Electron; Molecular and cellular biology; Molecular immunology; Mouse; Phenotype; Techniques; Thymidine - metabolism
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(01)00371-4

Vascular endothelium is an important site for a wide array of immunological processes such as inflammation, atherosclerosis and allograft rejection. Culture methods of mouse vascular endothelium would provide an important in vitro correlate to immunological murine in vivo models. We describe a simple method to culture mouse vascular endothelium from thoracic aorta. Our cultured cells express typical phenotypic (CD105, CD31, CD106), morphological and ultrastructural (intercellular junctions, Weibel–Palade bodies) markers of vascular endothelium. They also possess functional receptors for uptake and processing of acetylated low-density lipoproteins. The mouse vascular endothelium within our system expresses high levels of MHC class I and MHC class II after activation with IFN-γ. In addition, these cells express the accessory molecules CD80 and CD54, while they lack constitutive expression of CD86 and CD40, providing them the means to function as antigen presenting cells. Alloreactive CD4 + and CD8 + T lymphocytes demonstrate evidence of DNA synthesis after co-culture with activated vascular endothelium indicating their commitment to proliferation. In conclusion, we describe a simple culture system to isolate and grow mouse vascular endothelium, which provides a powerful tool to study biological interactions in vitro.