Establishment of Rat Lymphatic Endothelial Cell Line

• Published in: Microcirculation (New York, N.Y. 1994) Vol. 10; no. 2; pp. 127 - 131
• Main Authors: Mizuno, Risuke, Yokoyama, Yumiko, Ono, Nobuyuki, Ikomi, Fumitaka, Ohhashi, Toshio
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-04-2003
• Subjects: acetylated low-density lipoprotein; culture; F-actin; factor VIII-related antigen; lymphatic endothelial cells; rat
• ISSN: 1073-9688; 1549-8719
• DOI: 10.1038/sj.mn.7800181

ABSTRACT
Objective: The objective of the present study was to establish a rat lymphatic endothelial cell line and then to investigate the morphological and immunohistochemical properties of the cells.
Methods: The lymphatic endothelial cells of rat thoracic ducts were isolated enzymatically by trypsin digestion and were cultured in endothelium growth medium (EGM)‐2 in an atmosphere of low oxygen (5% O2, 5% CO2, and 90% N2) or high oxygen (21% O2, 5% CO2, and 74% N2).
Results: The number of the cells cultured in the low‐oxygen atmosphere was significantly larger than that obtained in the high‐oxygen atmosphere. The cultured cells in the low‐oxygen atmosphere showed a monolayer with uniform cobblestone appearance, suggesting the morphological properties of endothelial cells. Factor VIII‐related antigen and cell surface carbohydrates (i.e., D‐galactose α and D‐N‐acetylgalactosamine α) were found on the lymphatic cultured cells. The phagocytosis of 1,1‐diocadecyl1‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate‐labeled acetylated low‐density lipoprotein also was observed in the cultured cells. The cytoskeleton protein F‐actin was located on the plasma membrane of the cultured cells as circumferential thin bundles and in the cytoplasma as filamentous bundles.
Conclusions: The present study indicates that the choice of EGM‐2 as a culture medium and the hypoxic atmosphere (∼5%) enabled us to establish rat lymphatic endothelial cell line.