| Field | Specification |
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| Mfr No | |
| Species |
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.
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- Protein expression: Keratin 8, Keratin 18, Vimentin.
- Tumorigenic: Yes, in C57BL/6J mice. In the third week, tumors will develop which are approx. 5-6 mm in diameter.
- Mutational profile: p53mut, C:G -+ G:C transversion at codon 132 of mouse p53 exon 5, which corresponds to an amino acid change from cysteine to tryptophan.
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 25 to 30 hours
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 to 2 x 104 cells/cm2 during routine culture
- fluidRenewal: Every 3 to 4 days
- postThawRecovery: >90% of the cells recovered from the freezing process within 24 to 48 h
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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