Immortalized Human Umbilical Vein Endothelial Cell Line

Frequently Asked Questions

• What are the Immortalized Human Umbilical Vein Endothelial Cells?

  The Immortalized Human Umbilical Vein Endothelial Cells are isolated from the endothelial cells lining the umbilical vein of the human umbilical cord and have been immortalized using Lenti-SV40T Lentivirus. These cells are used primarily for cardiovascular disease research and exhibit endothelial morphology with adherent growth properties.

• What is the recommended culture medium for these cells?

  The base medium for these cells is F-12K medium. To make the complete growth medium, add 5 mL of a 10 mg/mL heparin solution, 50 mL fetal bovine serum, and 500 µL of 30 mg/mL endothelial cell growth supplement to 450 mL of the base medium. The cells should be incubated at 37°C, 5% CO2.

• How can ensure proper growth of these cells?

  To ensure optimal viability and growth, thaw the cells vial by gentle agitation in a 37°C water bath for approximately 2 minutes, being careful to avoid water contact with the cap and O-ring. Post-thaw, cells should be handled under strict aseptic conditions and transferred into a centrifuge tube containing complete culture medium for centrifugation. Resuspend the cell pellet in recommended medium and seed at a density of 2.0 × 10^4 to 4.0 × 10^4 viable cells/cm². Before adding the vial contents, pre-equilibrate the culture vessel containing the growth medium in an incubator to allow the medium to reach its normal pH (7.0 to 7.6).

• What steps are recommended for subculturing these cells?

  For subculturing, it’s essential to use a T-25 flask and initiate the process when cells are 80 to 90% confluent to manage cell detachment and viability. Begin by rinsing the cell layer with trypsin-EDTA to remove serum, which contains trypsin inhibitor. Add 1.0 mL of trypsin-EDTA and observe until cells detach, then neutralize the trypsin with complete growth medium. Gently pipette to aspirate cells, and seed new culture vessels at a recommended subcultivation ratio of 1:2 to 1:3. Maintain the cultures by changing the medium two to three times per week and avoid excessive alkalinity during recovery.

• Why these cells are suitable for the vascular research?

  Immortalized Human Umbilical Vein Endothelial Cells are derived from the endothelial layer of the human umbilical vein, making them particularly relevant for studies involving the vascular endothelium. Their endothelial morphology and ability to form an interface between circulating blood and the vessel wall allow researchers to effectively simulate and study endothelial function and pathology under controlled laboratory conditions. This includes the investigation of endothelial barrier function, interactions with circulating cells, and the secretion of factors that regulate blood flow and inflammation. These characteristics make them an indispensable tool in vascular biology and related research areas.