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The line contains human papillomavirus type 18 (HPV-18) DNA sequences, and expresses HPV-18 RNA. ATCC confirmed this cell line is positive for the presence of HPV viral DNA sequences via PCR.

Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 2.0 to 3.0 mL of complete growth medium and aspirate cells by gently pipetting
5. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

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Herz F, et al. Chromosome analysis and alkaline phosphatase of C41, a cell line of human cervical origin distinct from HeLa. Cancer Res. 37: 3209-3213, 1977. PubMed: 560251

Yee C, et al. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am. J. Pathol. 119: 361-366, 1985. PubMed: 2990217

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Auersperg N. Histogenetic behavior of tumors. II. Roles of cellular and environmental factors in the in vitro growth of carcinoma cells. J. Natl. Cancer Inst. 43: 175-190, 1969. PubMed: 5796382

Auersperg N. Histogenetic behavior of tumors. 3. Possible relationships to patterns of glycolysis. J. Natl. Cancer Inst. 48: 1589-1596, 1972. PubMed: 5056250