| 產(chǎn)品名稱 |
pgsD-677 |
| 商品貨號 |
B165506 |
| 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 |
heparan 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 heparan sulfate.
PgsD-677 cells do not produce heparan sulfate. They do produce chondroitin sulfate.
They lack both N-acetylglucosaminyltransferase and glucuronyltransferase activities that are required for synthesis of heparan 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.
- Remove and discard culture medium.
- 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.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- 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.
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| Cryopreservation |
Culture medium, 95%; DMSO, 5% |
| Name of Depositor |
JD Esko |
| Deposited As |
Cricetulus griseus |
| 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
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