pgsB-650 (ATCC® CRL-2243)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  / 

Permits and Restrictions

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Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Gender female
Storage Conditions liquid nitrogen vapor phase
Derivation
The cell line was derived from CHO-K1 cells (see ATCC CCL-61) treated with mutagen (ethylmethanesulfonate) and screened for mutants defective in proteoglycan synthesis.
Clinical Data
female
Comments
PgsB-650 is a Chinese hamster ovary cell mutant deficient in galactosyltransferase I (UDP-D-galactose:xylose-1,4-D-galactosyl transferase).

PgsB-650 cells have reduced levels of galactosyltransferase, and have a three-fold reduction in production of glycosaminoglycans relative to normal.
Complete Growth Medium The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium: fetal bovine 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 1:8
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor JD Esko
References

Esko JD, et al. Tumor formation dependent on proteoglycan biosynthesis. Science 241: 1092-1096, 1988. PubMed: 3137658

Esko JD, et al. Inhibition of chondroitin and heparan sulfate biosynthesis in Chinese hamster ovary cell mutants defective in galactosyltransferase I. J. Biol. Chem. 262: 12189-12195, 1987. PubMed: 2957376

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
Restrictions

This line is available subject to the following:

  1. The CHO cell mutant was deposited in the ATCC by Dr. Jeffrey D. Esko and is provided for research purposes only as a service to the research community. They are provided without warranty or merchantability of fitness for a particular purpose or any other warranty, express or implied. The cells are provided with the understanding that they will not be used for commercial purposes.
  2. All products derived from the cells or genetically altered forms of the cells cannot be commercialized without express permission from the University of Alabama at Birmingham. Commercial interests are the exclusive property of the University of Alabama at Birmingham.
  3. Any proposed commercial uses of these cells must first be negotiated with the Research Foundation, University of Alabama at Birmingham, Birmingham, AL 35294. Telephone: (205) 934-9911.
  4. In all papers reporting any use of these cells or derived products, a direct reference will be made to the original publication listed above.

References

Esko JD, et al. Tumor formation dependent on proteoglycan biosynthesis. Science 241: 1092-1096, 1988. PubMed: 3137658

Esko JD, et al. Inhibition of chondroitin and heparan sulfate biosynthesis in Chinese hamster ovary cell mutants defective in galactosyltransferase I. J. Biol. Chem. 262: 12189-12195, 1987. PubMed: 2957376