| Field | Specification |
|---|---|
| Mfr No | |
| Species |
The Hep-55.1c hepatoma cell line is derived from a mouse liver tumor, specifically from the C57BL/6J mouse strain. This cell line is characterized by its hepatocytic origin, confirmed through intermediate filament protein analysis. Hep-55.1c expresses simple keratins K8 and K18, which are typical of normal liver cells, as well as vimentin and keratin K19 to varying degrees. These protein patterns confirm the hepatocytic nature of the cell line and its classification as a hepatoma line.
The Hep-55.1c cell line displays a predominantly epithelial morphology, reflecting its origin from hepatocytes. This morphological phenotype is consistent with its protein expression profile. DNA fingerprint analysis of Hep-55.1c did not reveal any major structural abnormalities, indicating a degree of genomic stability. However, some changes in the relative intensities of specific bands were observed with increasing passage numbers, suggesting minor genomic variability over extended culture periods.
Despite the absence of detectable p53 mutations in the primary mouse liver tumors, aberrations were found in some hepatoma lines during in vitro propagation. The Hep-55.1c cell line was analyzed for mutations in the p53 and c-Ha-ras genes. The absence of detectable mutations in the p53 gene in this line during early passages suggests a stable genetic background. This cell line serves as a valuable model for studying hepatocellular carcinoma, providing insights into the cellular and molecular mechanisms underlying liver tumorigenesis.
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- Protein expression: Keratin 8, Keratin 18, Vimentin.
- Tumorigenic: Yes, in C57BL/6J mice
- Mutational profile: p53 wt
- 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
- 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.
- fluidRenewal: Every 3 to 5 days
- postThawRecovery: Start culture from cryovial at a cell density of 3 to 4 x 104 cells/cm2. The cells will recover within 24 to 48 hours.
- 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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