Culture and characterization of human umbilical vein endothelial cells

ABSTRACT

Endothelial cells, the lining cells of blood vessels, are involved in many physiological and pathophysiological processes, including haemostasis, vasoregulation, inflammation, angiogenesis, and the extravasation of fluids, macromolecules, hormones, and leucocytes. This work was carried out to investigate the isolation, culture and characterization of human umbilical vein endothelial cells [HUVECs] in vitro and also to improve the cell yield and growth of these cells in culture. The endothelial cells were isolated by collagenase digestion and were seeded in tissue culture flasks using Dulbecco's modified Eagles medium [DMEM]. This study showed that the HUVECs were attached completely after 24 hour and became confluent after 5 days in vitro. The HUVECs displayed cobblestone morphology at confluence with granular cytoplasm. Immunohistochemically, these cells were positively stained using CD34 monoclonal antibody. The cells produced from this technique grew well for extended periods and retained the normal characteristics of primary HUVECs, making them invaluable tools for researches