A modified protocol for the isolation and culture of human umbilical vein endothelial cells

The umbilical cord is a valuable source of foetal stem cells, progenitor cells, and early-stage developmental cells, including human umbilical vein endothelial cells (HUVECs). HUVECs are widely used as a model for endothelial biology and are increasingly being investigated for their regenerative potential. Efficient isolation of these cells from the umbilical vein is a critical first step for both research and therapeutic applications. To date, most published protocols utilise Collagenase A for isolation. In this study, we present a modified HUVEC isolation protocol that employs dispase, alongside refined tissue and cell culture handling practices. We characterised the isolated cells using established HUVEC markers CD31 and CD146, and demonstrated in situ detachment of the cells from the vessel wall through immunofluorescence imaging. Our method achieved a success rate exceeding 95.6 % across all umbilical cords processed. These findings highlight the protocol's potential for broad applicability across research settings, using readily accessible reagents and equipment.