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Hepatitis B virus (HBV) DNA was detected by Southern blot hybridization. HBV genomic RNA was not expressed.

Grossly, the original tumor was single nodular with perinodular extensions. Histologically, it was predominantly compact and minor trabecular type.

The cultured cells contain a single or double nucleus.

ATCC confirmed this cell line is positive for the presence of Hepatitis B viral DNA sequences.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C

Park JG, et al. Characterization of cell lines established from human hepatocellular carcinoma. Int. J. Cancer 62: 276-282, 1995. PubMed: 7543080