M1WT3 (ATCC® CRL-1985)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  / 

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Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Gender female
Storage Conditions liquid nitrogen vapor phase
Derivation
The M1WT3 cell line was derived by transfecting CHO-K1 cells (ATCC CCL-61) with the pSVL expression vector containing the gene for the rat m1 muscarinic acetylcholine receptor.

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

Receptor Expression
acetylcholine, muscarinic m1
Comments
M1WT3 cells are reported to express 800 fmol of receptor protein per mg of membrane protein.
Complete Growth Medium Ham's F12K medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate supplemented with 0.05 to 0.1 mg/ml G418, 90%; fetal bovine serum, 10%
Subculturing Volumes used in this protocol are for 75 cm2 flasks; 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 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.
  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 with 5 to 10 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 cell suspension to new culture vessels.
  6. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: At the time of subcultivation

Cryopreservation
Freeze medium: Complete growth medium, 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Name of Depositor CM Fraser
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
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