Technical information sheet for CloneticsTM HUVEC cells

Protocol for Angiogenesis assay using HUVEC cells

In this study, we have optimized this assay using Lonza's Primary HUVECs and EGM®-2 Media

HUVECs are a good primary endothelial cell type for many in vitro studies and also for several high-throughput screening applications. Consequently, Lonza offers, HUVEC-XL. These cells are expanded to passage-3 from pooled P1-HUVECs and packaged at 10 million cells per ampoule.

Human Umbilical Vein Endothelial Cells seeded on BME and the formation of endothelial tubes were monitored live with the CytoSMART™ Lux 10X System, the previous version of the CytoSMART™ 2 System. Time-lapse video was recorded for 22 hours at a frame interval of 1 frame every 5 minutes.

Time-Lapse Video of Human Umbilical Vein Endothelial Cell Angiogenic Tube Formation in Co-Culture with Dermal Fibroblasts. Primary Human Umbilical Vein Endothelial Cells (HUVECs) were co-cultured with primary Human Normal Dermal Fibroblast cells at a ratio of 1:3 HUVECs: fibroblasts and a total cell density of 40,000 cells/well in a 96-well plate. Co-cultures were maintained for 5 days with medium changes every 2-3 days in EBMTM-2 medium supplemented with a custom concentration of VEGF (313 pM) to optimize angiogenic tube formation in this setting. Cells were stained with CellTracker® Dye from ThermoFisher. Purple lines indicate angiogenic tube formation detected by the live cell imaging software. At each frame, the software quantifies angiogenesis by counting the number of network branch points (i.e., the number of branches in each network of tubes) per mm2 and the mean network length (i.e., the average of the total network length of tubes per mm2).

Lonza's Primary HUVECs are an excellent cell type to measure the effects of pharmacologically-important compounds on vial signaling pathways, and they provide increased data relevancy compared to cell lines.

Lonza, the market leader in primary cells, has expanded its human primary cell portfolio with an improved version of Human Umbilical Vein Endothelial Cells (HUVEC) cultured in the absence of additive vascular endothelial growth factor (VEGF).

Instructions for culturing Clonetics™ HIVEC-XL™Pooled Cells