Caki-2

Caki-2 is a human clear cell renal cell carcinoma (ccRCC) cell line that exhibits epithelial morphology and adheres during in vitro culture conditions. It serves as an essential preclinical model for the investigation of renal cancer mechanisms and therapeutic responses. The Caki-2 line is particularly notable for its resistance to certain chemotherapeutic agents; it displays decreased sensitivity to 5-fluorouracil and the multi-kinase inhibitor sorafenib, which targets VEGFRs 1-3, PDGFR-b, and Raf-1, in comparison to the Caki-1 cell line. This differential sensitivity is significant for studying drug resistance mechanisms and evaluating new therapeutic strategies in renal cell carcinoma.

The genetic background of Caki-2 cells includes a loss-of-function mutation in the von Hippel-Lindau (VHL) tumor suppressor protein, a hallmark of many ccRCCs that leads to the deregulation of hypoxia-inducible factors (HIFs) and contributes to tumorigenesis. The ability of Caki-2 cells to form tumors in immunocompromised mice makes them a valuable tool for in vivo studies of cancer growth and metastasis, providing insights into the tumor environment and potential therapeutic interventions. Their use extends to exploring the role of VHL in cancer progression and testing the efficacy of drugs targeting the HIF pathway and other associated signaling cascades in a controlled experimental setup.

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.