pgsE-606 (ATCC® CRL-2246)

Organism: Cricetulus griseus, hamster, Chinese  / 

Organism Cricetulus griseus, hamster, Chinese
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Gender female
Applications
PgsE-606 is a Chinese hamster ovary cell mutant partially deficient in heparin sulfate N sulfotransferase.
The cell line was derived from CHO-K1 cells (see ATCC CCL-61) treated with mutagen (ethylmethanesulfonate), and isolated by replica plating [35S]sulfate colony autoradiography.
PgsE-606 produce an under sulfated form of heparin sulfate.
Derivation
The cell line was derived from CHO-K1 cells (see ATCC CCL-61) treated with mutagen (ethylmethanesulfonate), and isolated by replica plating [35S]sulfate colony autoradiography.
Clinical Data
female
Comments
PgsE-606 is a Chinese hamster ovary cell mutant partially deficient in heparin sulfate N sulfotransferase.
The cell line was derived from CHO-K1 cells (see ATCC CCL-61) treated with mutagen (ethylmethanesulfonate), and isolated by replica plating [35S]sulfate colony autoradiography.
PgsE-606 produce an under sulfated form of heparin sulfate.
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
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Protocol: Volumes used in this protocol are for 75 sq cm flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  • Remove and discard culture medium.
  • Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  • 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 37C to facilitate dispersal.
  • Add 2.0 to 3.0 ml of complete growth medium and aspirate cells by gently pipetting
  • Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  • Incubate cultures at 37C.
Cryopreservation
Culture medium, 95%; DMSO, 5%
Name of Depositor JD Esko
References

Bame KJ, Esko JD. Undersulfated heparan sulfate in a Chinese hamster ovary cell mutant defective in heparan sulfate N-sulfotransferase. J. Biol. Chem. 264: 8059-8065, 1989. PubMed: 2524478

Bame KJ, et al. Biosynthesis of heparan sulfate. Coordination of polymer-modification reactions in a Chinese hamster ovary cell mutant defective in N-sulfotransferase. J. Biol. Chem. 266: 10287-10293, 1991. PubMed: 2037581

Bame KJ, et al. Coupling of N-deacetylation and N-sulfation in a Chinese hamster ovary cell mutant defective in heparan sulfate N-sulfotransferase. J. Biol. Chem. 266: 12461-12468, 1991. PubMed: 2061321

Basic Documentation
References

Bame KJ, Esko JD. Undersulfated heparan sulfate in a Chinese hamster ovary cell mutant defective in heparan sulfate N-sulfotransferase. J. Biol. Chem. 264: 8059-8065, 1989. PubMed: 2524478

Bame KJ, et al. Biosynthesis of heparan sulfate. Coordination of polymer-modification reactions in a Chinese hamster ovary cell mutant defective in N-sulfotransferase. J. Biol. Chem. 266: 10287-10293, 1991. PubMed: 2037581

Bame KJ, et al. Coupling of N-deacetylation and N-sulfation in a Chinese hamster ovary cell mutant defective in heparan sulfate N-sulfotransferase. J. Biol. Chem. 266: 12461-12468, 1991. PubMed: 2061321