title: Establishment of rat lymphatic endothelial cell line.
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% 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.
authors.affiliation: The First Department of Physiology, Shinshu University School of Medicine, Matsumoto, Japan.
authors.forename: Risuke
authors.initials: R
authors.lastname: Mizuno
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Yokoyama
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Ono
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Ikomi
authors.forename: Toshio
authors.initials: T
authors.lastname: Ohhashi
mesh_terms: 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
chemical_list: D000199:Actins; D014841:von Willebrand Factor; D010100:Oxygen