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clone 1-5c-4 [Wong-Kilbourne derivative (D) of Chang conjunctiva] (ATCC® CCL-20.2)

Organism: Homo sapiens, human  /  Tissue: epithelial  / 

Permits and Restrictions

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Organism Homo sapiens, human
Tissue epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain human papilloma virus (HPV-18)

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Storage Conditions liquid nitrogen vapor phase
Derivation
This line was originally thought to be derived from normal conjunctiva, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination.
HeLa Markers Y
Genes Expressed
keratin
Cellular Products
keratin
Comments
The cells are positive for keratin by immunoperoxidase staining.

This cell line contains HeLa marker chromosomes and has type A G6PD (see Nature 259:211-213, 1976 and In Vitro 14:469-475, 1978).
Note: Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination

Complete Growth Medium The base medium for this cell line is Medium 199 containing Earle's BSS and 2.2 g/L sodium bicarbonate. To make the complete growth medium, add the following component to the base medium: bovine calf serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; 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 0.25% (w/v) Trypsin-0.53mM 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 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.

Subcultivation Ratio: 1:4 to 1:10
Medium Renewal: Every 2 to 3 days

Note:For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

Cryopreservation
Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC® Catalog No. 4-X.

Culture Conditions

Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%

STR Profile
Amelogenin: X
CSF1PO: 9,10
D13S317: 12,13.3
D16S539: 9,10
D5S818: 11,12
D7S820: 8,12
TH01: 7
TPOX: 8,12
vWA: 16,18
Isoenzymes
G6PD, A
Name of Depositor ED Kilbourne
Deposited As Homo sapiens
References

Wong SC, Kilbourne ED. Changing viral susceptibility of a human cell line in continuous cultivation. I. Production of infective virus in a variant of the Chang conjunctival cell following infection with swine or N-WS influenza viruses. J. Exp. Med. 113: 95-110, 1961. PubMed: 13786478

Chang RS. Continuous subcultivation of epithelial-like cells from normal human tissues. Proc. Soc. Exp. Biol. Med. 87: 440-443, 1954. PubMed: 13237268

. . Virology 26: 478, 1965.

Gomez-Duarte OG, et al. Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invastion of HeLa cells. Infect. Immun. 65: 3857-3866, 1997. PubMed: 9284164

St. Geme JW, et al. Characterization of the genetic locus encoding Haemophilus influenzae type b surface fibrils. J. Bacteriol. 178: 6281-6287, 1996. PubMed: 8892830

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Wong SC, Kilbourne ED. Changing viral susceptibility of a human cell line in continuous cultivation. I. Production of infective virus in a variant of the Chang conjunctival cell following infection with swine or N-WS influenza viruses. J. Exp. Med. 113: 95-110, 1961. PubMed: 13786478

Chang RS. Continuous subcultivation of epithelial-like cells from normal human tissues. Proc. Soc. Exp. Biol. Med. 87: 440-443, 1954. PubMed: 13237268

. . Virology 26: 478, 1965.

Gomez-Duarte OG, et al. Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invastion of HeLa cells. Infect. Immun. 65: 3857-3866, 1997. PubMed: 9284164

St. Geme JW, et al. Characterization of the genetic locus encoding Haemophilus influenzae type b surface fibrils. J. Bacteriol. 178: 6281-6287, 1996. PubMed: 8892830

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.