HEK293-VEGF-TM

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HEK293-VEGF-TM cells are human embryonic kidney 293 (HEK293) cells engineered to stably express a membrane-tethered form of vascular endothelial growth factor (VEGF), commonly designed to present VEGF at the cell surface through fusion with a transmembrane domain. Unlike soluble VEGF isoforms that are secreted into the extracellular environment, VEGF-TM constructs enable localized and sustained presentation of VEGF ligands on the plasma membrane, facilitating controlled investigation of VEGF receptor interactions and cell-cell signaling mechanisms. These engineered models are useful for studying angiogenic signaling pathways mediated primarily through VEGFR1 (FLT1) and VEGFR2 (KDR), which regulate endothelial proliferation, migration, vascular permeability, and neovascularization.

HEK293-VEGF-TM cells are widely used in angiogenesis research and therapeutic development for characterization of VEGF-targeted antibodies, receptor traps, anti-angiogenic biologics, and engineered immune cell therapies. The membrane-anchored VEGF presentation enables quantitative assessment of receptor binding, ligand accessibility, antibody blockade, receptor clustering, and cell-contact-dependent signaling events. These cells are particularly valuable in flow cytometry-based binding assays, co-culture systems, reporter assays, and high-throughput screening platforms evaluating VEGF/VEGFR pathway inhibition. In addition, HEK293-VEGF-TM models can support studies examining synapse formation and target recognition by VEGF-directed CAR-T cells or bispecific antibody platforms.

37°C, 5% CO₂, humidified atmosphere.

After thawing, split the cells at a ratio of 1:2 to 1:3 in T25 flasks and allow the cells to recover from the freezing process and to adhere for at least 24 hours.

For best attachment and viability after thawing the cells, we recommend using Collagen-coated flasks or plates for the initial seeding after cryo-recovery. Collagen coating is not required for subsequent routine culture of the cells.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.