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L Wnt-5A (ATCC® CRL-2814™)

Organism: Mus musculus, mouse  /  Tissue: subcutaneous connective tissue; areolar and adipose  / 

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Organism Mus musculus, mouse
Tissue subcutaneous connective tissue; areolar and adipose
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Applications
They are presently the best source for production of Wnt-5A conditioned medium. Since the conditioned medium contains other factors besides the Wnt-5A protein, it is necessary to control any experiments involving the Wnt-5A conditioned medium with control conditioned medium from the parental cell line (ATCC CRL-2648).
Storage Conditions liquid nitrogen vapor phase
Images
Derivation L-M(TK-) cells (ATCC CCL-1.3) were transfected with a Wnt-5A expression vector and stable clones were selected in medium containing G418.
Comments

The Wnt-5A gene encodes a secreted glyoprotein with a variety of signaling effects. Wnt proteins bind to receptors of the Frizzled family on the cell surface. Wnt genes control many of the patterning and growth events during embryonic development. 

The cells secrete biologically active Wnt-5A protein. 
Complete Growth Medium The base medium for this cell line is ATCC Hybri-Care Medium, Catalog No. 46-X. Hybri-Care Medium is supplied as a powder and should be reconstituted in 1 L cell-culture-grade water and supplemented with 1.5 g/L sodium bicarbonate. 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-0.53 mM EDTA solution to remove all traces of serum that 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 pipetting gently.
  5. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 6 x 103 to 8 x 103 viable cells/cm2 is recommended.
  6. Incubate culture vessels at 37°C. We recommend that you maintain cultures at a cell concentration between 5 x 104 and 3 x 105 cells/cm2.

Subcultivation Ratio: 1:4 to 1:6
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
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Population Doubling Time 36 hrs
Name of Depositor R Nusse
References

Chen W, et al. Dishevelled 2 recruits beta-arrestin 2 to mediate Wnt5A-stimulated endocytosis of Frizzled 4. Science 301: 1391-1339, 2003. PubMed: 12958364

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
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
MSDS
MSDS
Other Documentation
Other Document
Cell Micrograph
References

Chen W, et al. Dishevelled 2 recruits beta-arrestin 2 to mediate Wnt5A-stimulated endocytosis of Frizzled 4. Science 301: 1391-1339, 2003. PubMed: 12958364