JHH-2

JHH-2 is a human hepatocellular carcinoma (HCC) cell line established from an ascites metastasis of a 56-year-old Japanese male patient. The line grows as an adherent monolayer with spindle-shaped cell morphology. JHH-2 harbors a TP53 p.His83Tyr heterozygous mutation affecting the transactivation domain of p53, which impairs wild-type p53 transcriptional activity and is associated with reduced apoptotic responses to DNA damage. This mutation, combined with the metastatic ascites origin of the line, makes JHH-2 a relevant model for studying advanced, metastatic HCC biology.

JHH-2 is applicable in hepatocellular carcinoma research, including studies of HCC cell invasion and migration, TP53 mutation-associated phenotypes, resistance to sorafenib and other targeted agents, and evaluation of novel therapeutic strategies against advanced HCC. The ascites origin suggests an aggressive, metastatic phenotype, making JHH-2 suitable for studies of HCC dissemination and the biology of liver cancer with acquired p53 dysfunction. It is also used as a xenograft model in immunocompromised hosts.

JHH-2 is maintained as an adherent culture in Williams' E medium supplemented with 10% FBS (originally 10%, subsequently 2% FCS after 5 years; alternative: MEM + 10% FBS). Cells are subcultured using 0.02% EDTA + 0.25% trypsin. Recommended seeding density is 1 × 10⁵ cells/cm². Doubling time is approximately 33–150 hours depending on conditions.

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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

7. Follow the procedure described under Post-Thaw Recovery

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.