CHON-002 (ATCC® CRL-2847)

Organism: Homo sapiens, human  /  Cell Type: Fibroblast immortalized with hTERT  /  Tissue: Long bone; cartilage  /  Disease: Normal

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Organism Homo sapiens, human
Tissue Long bone; cartilage
Cell Type Fibroblast immortalized with hTERT
Product Format frozen
Morphology Fibroblast-like
Culture Properties Adherent
Biosafety Level 2 [Cells contain SV40 viral DNA sequences]
Disease Normal
Age Fetal, 18 weeks
Gender Female
Storage Conditions Liquid nitrogen vapor phase
Karyotype This is a diploid cell line of female origin. Overall, the karyology is stable with a modal chromosome number of 46 in 87% of the examined cells and a low rate of polypoidy. No consistent structural chromosomal aberrations were found in any of the cells examined.
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Derivation
The chondrocyte cell line, CHON-002, was derived from the long bones of an 18-week female fetus. The primary cells were infected by the defective retrovirus containing hTERT gene under G418 selection. The defective retrovirus was collected from the supernatant of the packaging cell line, PT67 transfected with the pLXSN vector containing hTERT gene.
Antigen Expression This cell line expresses the following (Homo sapiens) proteins: HLA-B81, HLA-B58, HLA-Cw6, HLA-Cw8, HLA-DR7, HLA-DR12, HLA-DR52, HLA-DQ2, HLA-DQ5.
Genes Expressed
Positive for Collagen type I and aggrecan (RT-PCR). Negative for Collagen type II (RT-PCR).
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • 0.1mg/ml G-418
  • 10% heat-inactivated fetal bovine serum

Subculturing
Volumes are given for T-75 flasks; proportionally reduce or increase the amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Add 2.0-3.0 mL of 0.05% Trypsin-0.53mM EDTA solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 10 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 to detatch may be placed at 37°C to facilitate dispersal.
  3. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  4. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 15 minutes.
  5. Discard supernatant and resuspend cells in fresh growth medium.
  6. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 5 X 103 to 7 X 103 viable cells/cm2 is recommended. Subculture when cell concentration reaches between 2 x 104 and 3 x 104 cells/cm2.
  7. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Aminal Cells: A Manual of Basic Techniques by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
Cryopreservation
Freeze medium: 90% heat-inactivated fetal bovine serum; 10% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 10,11
D13S317: 12,14
D16S539: 11,13
D5S818: 11,12
D7S820: 12
THO1: 6,7
TPOX: 8,11
vWA: 16,17
Population Doubling Level (PDL) As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation. We have also compared its karyotype, telomerase expression level, growth rate, morphology and tissue-specific markers when first recovered from cryopreservation with that of cells at 10+ population doublings to ensure that there is no change in these parameters and that the cells are capable of extended proliferation.
Name of Depositor ATCC
Year of Origin December 20, 2001
References

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

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.

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.
  • License agreement required for commercial customer uses.
  • This material is distributed for research purposes only. A signed addendum to the ATCC Material Transfer Agreement must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

This material is subject to claims under U.S. Patent Nos. 6,261,836 and 6,337,200, other pending patent applications, and foreign counterparts thereof. It is required that either the Addendum for Commercial and For-Profit Organizations or the Addendum for Noncommercial and Academic Organizations, as appropriate, be signed and returned to ATCC before shipment.

References

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

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.