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This line was originally thought to be derived from normal liver tissue, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination.
The cells are positive for keratin by immunoperoxidase staining.
NOTE: Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination
Propagation:
ATCC complete growth medium: Minimum essential medium (Eagle) in Earle's BSS with non-essential amino acids and 1 mM sodium pyruvate, 90%; fetal bovine serum, 10% Temperature: 37.0°C
Subculturing:
Protocol: Remove medium, add fresh 0.25% trypsin, 0.02% EDTA solution, rinse and remove. Allow flask to sit at room temperature (or at 37C) until cells detach (approximately 10 minutes). Add fresh medium, aspirate and dispense into new flasks. Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Preservation:
Complete growth medium, 95%; DMSO, 5%
References:
22213: Chang RS. Continuous subcultivation of epithelial-like cells from normal human tissues. Proc. Soc. Exp. Biol. Med. 87: 440-443, 1954. PubMed: 13237268 26122: Murphy WH, Landau BJ. Clonal variation and interaction of cells with viruses. Natl. Cancer Inst. Monogr. 7: 249-271, 1962. PubMed: 14477443 32352: Benn J, et al. Hepatitis B virus HBx protein induces transcription factor AP-1 by activation of extracellular signal-regulated and c-Jun N-terminal mitogen-activated protein kinases. J. Virol. 70: 4978-4985, 1996. PubMed: 8764004
