pgsD-677 (ATCC® CRL-2244)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  /  Disease: heparin sulfate deficient

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
Disease heparin sulfate deficient
Gender female
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
PgsD-677 is a Chinese hamster ovary cell mutant deficient in the polymerization of heparin sulfate.

PgsD-677 cells do not produce heparin sulfate. They do produce chondroitin sulfate.

They lack both N-acetylglucosaminyltransferase and glucuronyltransferase activities that are required for synthesis 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 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-0.53 mM EDTA solution to remove all traces of serum that 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

Subculture Ratio: 1:4 to 1:8
Medium Renewal: 2 to 3 times a week.
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
Culture medium, 95%; DMSO, 5%
Name of Depositor JD Esko
References

Lidholt K, et al. A single mutation affects both N-acetylglucosaminyltransferase and glucuronosyltransferase activities in a Chinese hamster ovary cell mutant defective in heparan sulfate biosynthesis. Proc. Natl. Acad. Sci. USA 89: 2267-2271, 1992. PubMed: 1532254

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

Basic Documentation
Restrictions

NOTE: 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

Lidholt K, et al. A single mutation affects both N-acetylglucosaminyltransferase and glucuronosyltransferase activities in a Chinese hamster ovary cell mutant defective in heparan sulfate biosynthesis. Proc. Natl. Acad. Sci. USA 89: 2267-2271, 1992. PubMed: 1532254

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