Hep-56.1D

The Hep-56.1D hepatoma cell line is derived from a mouse liver tumor, specifically from the C57BL/6J mouse strain. This cell line is characterized by a notable mutation in the p53 gene, identified at different passages during in vitro propagation. Specifically, Hep-56.1D exhibits a C:G to G:C transversion at codon 132 of exon 5, resulting in an amino acid change from cysteine to tryptophan. This mutation was detected at passage number 17, suggesting a selective growth advantage conferred by the mutation, leading to its predominance in the cell population.

The Hep-56.1D cell line displays a predominantly epithelial morphology, reflecting its hepatocytic origin. This is consistent with its intermediate filament protein profile, which includes the simple keratins K8 and K18, as well as vimentin and keratin K19 to varying degrees. The presence of these proteins confirms the hepatocytic nature of the cell line and its classification as a hepatoma line.

Further analysis of Hep-56.1D using DNA fingerprinting did not reveal any major structural abnormalities, although some changes in the relative intensities of specific bands were observed with increasing passage numbers. This indicates genomic stability with some degree of variability over extended culture periods. The p53 mutation analysis and intermediate filament protein expression patterns together establish Hep-56.1D as a valuable model for studying hepatocellular carcinoma and the role of p53 mutations in liver tumorigenesis.

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.