KYSE-30

KYSE-30 is a well-differentiated human esophageal squamous cell carcinoma (ESCC) cell line derived from a primary tumor in an adult patient. As part of the KYSE series, this cell line was established to study the molecular and cellular characteristics of esophageal cancer. KYSE-30 cells are notable for their rapid proliferation, with a doubling time of 20.8 hours, making them a robust model for in vitro cancer research. These cells grow predominantly as adherent monolayers, displaying a characteristic polygonal shape and uniform appearance under phase-contrast microscopy. Their growth pattern is typical of epithelial-derived cancer cells, forming tightly packed colonies with a tendency to pile up in a disorganized manner, reflecting the invasive nature of the tumor from which they were derived.

Genetically, KYSE-30 is significant for its alterations in key tumor suppressor genes. The cell line exhibits a wild-type configuration for the p16 (INK4a) and p15 (INK4b) genes, but it carries a notable point mutation in the p16 gene that results in a premature stop codon, leading to a truncated, non-functional protein. This mutation likely contributes to the loss of cell cycle control, promoting the unchecked proliferation characteristic of cancer cells. The retention of the wild-type p15 gene, however, suggests that p16 gene alterations play a more critical role in the oncogenesis of KYSE-30, which may be relevant in studies focusing on the differential roles of these genes in cancer.

KYSE-30 is tumorigenic, as demonstrated by its ability to form tumors when injected into athymic nude mice, making it an excellent model for in vivo studies of ESCC. The histological examination of tumors formed by KYSE-30 cells shows characteristics similar to the original squamous cell carcinoma, providing a faithful representation of the disease. This cell line is invaluable for research into the mechanisms of tumorigenesis, the genetic and epigenetic changes driving esophageal cancer, and the development of targeted therapies, although it is not suitable for therapeutic or in vivo applications.

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.