C 211 (ATCC® CCL-123)

Organism: Homo sapiens, human  /  Cell Type: fibroblast  /  Tissue: skin  /  Disease: Cri du Chat syndrome

Organism Homo sapiens, human
Tissue skin
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease Cri du Chat syndrome
Age 11 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen storage temperature
Karyotype model number = 46; range = 39 to 95.
Karyotype is that of a normal human diploid male with one 4 or 5 chromosome with a deleted short arm in all cells. The four cells with 47 chromosomes all had an extra G group chromosome.
Clinical Data
male
Caucasian
11 years old
Comments
The line has a life expectancy of approximately 21 passages.
Complete Growth Medium McCoy’s 5a medium (modified) with 1.5 mM L-glutamine adjusted to contain 2.2 g/L sodium bicarbonate, 80%; fetal bovine serum, 20%
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: 1:2 to 1:3
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%; CO2, 5%
STR Profile
Amelogenin: X,Y
CSF1PO: 11,12
D13S317: 12,13
D16S539: 9,11
D5S818: 9,12
D7S820: 10,11
THO1: 9.3
TPOX: 8
vWA: 16
Isoenzymes
G6PD, B
Name of Depositor WR Breg
Passage History
The line has a life expectancy of at least 30 passages.
References

Miller OJ, et al. Alternative DNA replication patterns associated with long arm length of chromosomes 4 and 5 in the cri du chat syndrome. Cytogenetics 5: 137-151, 1966. PubMed: 5921496

Warburton D, et al. Distinction between chromosome 4 and chromosome 5 by replication pattern and length of long and short arms. Am. J. Hum. Genet. 19: 399-415, 1967. PubMed: 6026933

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Basic Documentation
References

Miller OJ, et al. Alternative DNA replication patterns associated with long arm length of chromosomes 4 and 5 in the cri du chat syndrome. Cytogenetics 5: 137-151, 1966. PubMed: 5921496

Warburton D, et al. Distinction between chromosome 4 and chromosome 5 by replication pattern and length of long and short arms. Am. J. Hum. Genet. 19: 399-415, 1967. PubMed: 6026933

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm