Tissue culture of human and canine thoracic duct endothelium

Endothelial cells from the canine or human thoracic duct were harvested using 0.2% collagenase digestion and grown in Media 199 supplemented with fetal bovine serum. The canine endothelial cells grew to confluence (4.4 to 12 X 10(4) cells/cm2) in 6 to 10 d; doubling times ranged from 1.5 to 2.8 d. There was a minimum critical density for cell growth between 5000 and 10 000 cells/cm2. The canine endothelial cells have been maintained in culture for periods up to 11 mo. The human thoracic duct endothelial cells are more difficult to grow and maintain. Endothelial cells were isolated from 5 out of 35 human thoracic ducts and grew for periods of up to 2 wk before degenerating. Both human and canine endothelial cells were Factor VIII positive. It has thus been demonstrated that it is possible to grow canine and, less easily, human thoracic duct endothelium in tissue culture.