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M3WT5

規格:

價格:

編號:B165053

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

規格:

凍干粉

斜面

甘油

平板

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產品名稱

M3WT5

商品貨號

B165053

Organism

Cricetulus griseus, hamster, Chinese

Tissue

ovary

培養基

Product Format

frozen

Morphology

epithelial

Culture Properties

adherent

Biosafety Level

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.

Gender

female

Applications

The M3WT5 cell line was derived by transfecting CHO-K1 cells (ATCC CCL-61) with the pSVL expression vector containing the gene for the rat m3 muscarinic acetylcholine receptor.

Storage Conditions

liquid nitrogen vapor phase

Derivation

The M3WT5 cell line was derived by transfecting CHO-K1 cells (ATCC CCL-61) with the pSVL expression vector containing the gene for the rat m3 muscarinic acetylcholine receptor.

Clinical Data

female

Receptor Expression

acetylcholine, muscarinic m3

Comments

The M3WT5 cell line was derived by transfecting CHO-K1 cells (ATCC CCL-61) with the pSVL expression vector containing the gene for the rat m3 muscarinic acetylcholine receptor.

The resulting cells were co-transfected with pMSVNeo and selected for growth in medium containing 0.5 mg/ml G418.

M3WT5 cells are reported to express 690 fmol of receptor protein per mg of membrane protein.

Complete Growth Medium

Ham's F12 medium with 0.05 to 0.1 mg/ml G418, 90%; fetal bovine serum, 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 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 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: 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 13 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO. 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

CM Fraser

Deposited As

Cricetulus griseus

References

Buck MA, Fraser CM. Muscarinic acetylcholine receptor subtypes which selectively couple to phospholipase C: pharmacological and biochemical properties. Biochem. Biophys. Res. Commun. 173: 666-672, 1990. PubMed: 2175603

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.

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