HUH-1

The HuH-1 cell line is derived from a human hepatocellular carcinoma (HCC) and is used extensively in liver cancer research. This cell line has shown the ability to produce and secrete various liver-specific proteins, including those typically found in human plasma. Notably, HuH-1 cells, like other human hepatoma cell lines, can express and release hepatitis B surface antigen (HBsAg) in culture, providing a model for studying hepatitis B virus-related HCC mechanisms. The ability to produce HBsAg in culture suggests that HuH-1 cells retain differentiated functions similar to those of liver cells in vivo.

HuH-1 cells are also instrumental in the investigation of regulatory mechanisms in liver cancer, particularly those involving microRNAs and their targets. For example, studies have demonstrated that miR-182 upregulation can enhance the metastatic potential of HCC cells, including HuH-1, by downregulating metastasis suppressor 1 (MTSS1). This downregulation promotes invasive behavior and highlights the role of specific microRNAs in modulating gene expression and influencing cancer cell behavior. Such insights are valuable for understanding the progression of HCC and developing targeted therapeutic strategies.

The HuH-1 cell line's utility extends to research on the tumor microenvironment and its interactions with viral elements, making it a suitable model for exploring the intricate relationships between oncogenes, suppressor genes, and viral proteins. This cell line aids in elucidating the multifaceted pathways of hepatocarcinogenesis and the development of anti-cancer therapies tailored to liver cancer's molecular underpinnings.

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.