Establishment of human mucosal microvascular endothelial cells from inferior turbinate in culture

Abstract

Human microvascular endothelial cells were isolated and cultured from the mucosa of inferior turbinates. Using dish-coated collagen and a medium composed of a 1:1 mixture of sarcoma 180-conditioned medium and Dulbecco's modified Eagle's medium (containing 10% fetal bovine serum and 75 μg/mL endothelial cell growth factors prepared from bovine pituitary glands), these cells grew rapidly to confluence and survived serial passages until the 16th population doubling level. The cells were identified as endothelial cells by their morphology, immunostaining of factor VIII antigen, and cytochemical staining with Ulex europeus agglutinin. Furthermore, Weibel-Palade bodies and numerous pinocytotic vesicles were confirmed by electron microscopy. Proliferation experiments demonstrated the need for either endothelial cell growth factor or tumor-conditioned medium. An exogenous matrix was also required for these cells in tissue culture. A tubule-like morphology appeared in the original monolayer of human microvascular endothelial cells after 1 month in the same plate, indicating that these cells have the ability to form tubules in the presence of sarcoma 180-conditioned medium.