Establishment of rat lymphatic endothelial cell line.
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% O(2), 5% CO(2), and 90% N(2)) or high oxygen (21% O(2), 5% CO(2), and 74% N(2)).


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 alpha and D-N-acetylgalactosamine alpha) were found on the lymphatic cultured cells. The phagocytosis of 1,1-diocadecyl1-3,3,3',3'-tetramethylindo-carbocyanine 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 ( approximately 5%) enabled us to establish rat lymphatic endothelial cell line.
Mizuno
Risuke
R
The First Department of Physiology, Shinshu University School of Medicine, Matsumoto, Japan.
Yokoyama
Yumiko
Y
Ono
Nobuyuki
N
Ikomi
Fumitaka
F
Ohhashi
Toshio
T
D000199:Actins / Q000648:ultrastructure; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D048429:Cell Size; D042783:Endothelial Cells; D004729:Endothelium, Lymphatic / Q000166:cytology*; D007150:Immunohistochemistry; D008297:Male; D010100:Oxygen / Q000494:pharmacology; D010587:Phagocytosis; D051381:Rats; D017208:Rats, Wistar; D013897:Thoracic Duct / Q000166:cytology; D014841:von Willebrand Factor / Q000032:analysis
D000199:Actins; D014841:von Willebrand Factor; D010100:Oxygen