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Cells contain Human papillomavirus (HPV) sequences
Cells contain SV40 sequences
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The cells do not undergo growth arrest in cell culture due to exogenous expression of the telomerase and E6/E7 genes. CuFi-1 cells are homozygous for the delta F508 cystic fibrosis-causing mutation (delta F508/delta F508).
Another hTERT-immortalized cell line, derived from normal HAE is also available as ATCC CRL-4011 (NuLi-1). Both cell lines, when seeded on semipermeable filters and grown at the air-liquid interface, are capable of forming polarized differentiated epithelia that exhibit transepithelial resistance and maintain the ion channel physiology expected of each genotype.
Note: The culture flasks should be pre-coated with 60 μg/mL solution of Human Placental Collagen Type IV. (Sigma Cat. No. C-7521) at least 18 hours in advance then air-dried and rinsed 2-3 times with Dulbeccos Phosphate Buffered Saline.
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
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Halbert CL, et al. The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65: 473-478, 1991. PubMed: 1845902
Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332
Zabner J, et al. Development of cystic fibrosis and noncystic fibrosis airway cell lines . Am. J. Physiol. Lung Cell Mol Physiol. 284: L844-L854, 2003. PubMed: 12676769
Kiyono T, et al. Both Rb/p161NK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature 396: 84-88, 1998. PubMed: 9817205
Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13.