CHO-NECTIN4

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CHO-NECTIN4 cells are a stable recombinant Chinese hamster ovary (CHO) cell line engineered to express human Nectin-4 (also known as PVRL4 or Poliovirus receptor-related protein 4), a type I transmembrane protein belonging to the nectin family of cell adhesion molecules. Nectin-4 is a well-established tumor-associated antigen that is overexpressed in multiple solid tumor types, including urothelial bladder carcinoma, breast cancer, non-small cell lung cancer, and pancreatic cancer, making it a clinically validated target for antibody-drug conjugates (ADCs) and other targeted immunotherapeutics. The ADC enfortumab vedotin, which targets Nectin-4, is approved for treatment of urothelial carcinoma, underscoring the therapeutic relevance of this antigen.

CHO-NECTIN4 cells are widely used for the development and characterization of Nectin-4-directed antibodies, ADCs, bispecific antibodies, and CAR-T cell therapies. The stable recombinant expression system supports quantitative binding assays, ADCC/CDC cytotoxicity evaluations, receptor internalization studies, and high-throughput antibody screening by flow cytometry. The CHO background provides low endogenous expression of most human surface antigens, ensuring that observed signals are attributable to the stably expressed Nectin-4 transgene. This cell line is validated for use in drug discovery workflows, preclinical candidate selection, and mechanistic studies of Nectin-4 receptor biology.

For adherent cultures: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)

For suspension cultures: CHO Growth Medium A (from InSCREENeX; InSCREENeX catalog number INS-ME-1039)

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 adherent cultures) for at least 24 hours.

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.