CHO/dhFr- 倉(cāng)鼠卵巢細(xì)胞,二氫葉酸還原酶缺陷
ATCC® Number: CRL-9096?
Designations: CHO/dhFr- [CHO duk-, NOTE: dhfr refers to dihydrofolate reductase]
Depositors: Hoffmann-La Roche Ltd.
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Cricetulus griseus (hamster, Chinese)
Morphology: epithelial
Source: Organ: ovary
Disease: normal
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Applications: transfection host (Roche FuGENE® Transfection Reagents)
Gender: female
Propagation: ATCC complete growth medium: Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and supplemented with 0.1 mM hypoxanthine, 0.016 mM thymidine, 0.002mM Methotrexate (Amethopterin) and 10% fetal bovine serum
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Growth Conditions: The cells are deficient in dihydrofolate reductase.They should die in the absence of HT ( Hypoxanthine-Thymidine). Methotrexate (Amethopterin) is added to the cell culture medium to prevent growth of revertant cells with a low resistance to the drug.
Subculturing: Protocol:
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.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2005
recommended serum:ATCC 30-2020
Medium additive:ATCC 71-X
References: 26539: Shulman T, et al. An antibody reactive with domain 4 of the platelet-derived growth factor beta receptor allows BB binding while inhibiting proliferation by impairing receptor dimerization. J. Biol. Chem. 272: 17400-17404, 1997. PubMed: 9211881
30382: Morel-Kopp MC, et al. A three amino acid deletion in glycoprotein IIIa is responsible for type I glanzmann's thrombasthenia: importance of residues I1e325Pro326Gly327 for beta 3 integrin subunit association. Blood 90: 669-677, 1997. PubMed: 9226167
30522: Libyh MT, et al. A recombinant human scFv anti-Rh(D) antibody with multiple valences using a C-terminal fragment of C4-binding protein. Blood 90: 3978-3983, 1997. PubMed: 9354666
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