KYSE180

The KYSE180 cell line is a human esophageal squamous cell carcinoma (ESCC) model derived from a primary tumor. This cell line is classified as moderately differentiated and exhibits a range of genetic alterations characteristic of ESCC, including mutations in the NRF2 (NFE2L2) signaling pathway. Specifically, KYSE180 cells harbor the NRF2 D77V mutation, which contributes to the hyperactivation of NRF2, a transcription factor involved in oxidative stress response and tumor progression.

KYSE180 cells are widely used to study the molecular mechanisms of ESCC, including chemoresistance and oncogenic signaling. Elevated NRF2 activity in KYSE180 cells has been linked to resistance to chemotherapy and radiotherapy, as NRF2 enhances cellular antioxidant defenses and metabolic adaptations. Recent research has identified compounds like pyrimethamine that inhibit NRF2 signaling, resulting in reduced proliferation and tumorigenicity of KYSE180 cells. Such studies highlight the therapeutic potential of targeting NRF2 in NRF2-high cancers.

Additionally, KYSE180 cells have been utilized to evaluate the roles of the aldo-keto reductase (AKR) family of enzymes, particularly AKR1C1 and AKR1C2, in drug metabolism and chemoprevention. The expression of these enzymes is upregulated in KYSE180 cells, enhancing sensitivity to agents such as ethyl-3,4-dihydroxybenzoate (EDHB). This compound induces apoptosis and autophagy, demonstrating the cell line's utility in exploring drug responses and cellular stress mechanisms in ESCC.

37°C, 5% CO₂, humidified atmosphere.

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.