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pgsB-650

货号 TS211113
中文名称 null
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产品名称: pgsB-650
商品货号: TS211113
Organism: Cricetulus griseus, hamster, Chinese
Tissue: ovary
Product Format: frozen
Morphology: epithelial
Culture Properties: adherent
Biosafety Level: 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease: galactosyltransferase I deficient
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.xa0 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
Deposited As: Cricetulus griseus
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

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.