KYSE520

The KYSE520 cell line is a human esophageal squamous cell carcinoma (ESCC) model derived from a primary tumor. It is moderately differentiated and has been instrumental in investigating epithelial-mesenchymal plasticity (EMP) in esophageal cancer. KYSE520 cells exhibit heterogeneity, consisting of both epithelial-like (CD44v+) and mesenchymal-like (CD44v−) subpopulations. These two populations are capable of interconversion, reflecting a dynamic EMP process. This property makes KYSE520 an excellent model for studying cancer stem cell traits and chemoresistance mechanisms in ESCC.

Genetically, KYSE520 cells display notable epigenetic regulation. The promoter region of the JAM3 gene, a tumor suppressor, is unmethylated in these cells, allowing its expression. JAM3 plays a role in regulating cell proliferation, migration, and invasion through Wnt/β-catenin signaling. The maintenance of JAM3 expression in KYSE520 has been linked to the suppression of aggressive cancer phenotypes.

In therapeutic research, KYSE520 cells have been used to explore the role of fibroblast growth factor receptor-like 1 (FGFRL1). Studies have shown that FGFRL1-deficient KYSE520 cells exhibit reduced tumor growth and motility, alongside a decrease in matrix metalloproteinase-1 (MMP-1) and fibroblast growth factor binding protein 1 (FGFBP1) expression. These findings underscore the importance of FGFRL1 in tumorigenesis and suggest potential therapeutic targets. Additionally, the EMP dynamics and associated molecular pathways in KYSE520 cells provide insights into ESCC progression and resistance mechanisms, contributing to the development of targeted treatments.

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.