W-20-17 [W-20 clone 17] (ATCC® CRL-2623)

Organism: Mus musculus, mouse  /  Tissue: bone marrow; stroma  / 

Permits and Restrictions

View Permits

Organism Mus musculus, mouse
Tissue bone marrow; stroma
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Applications
This cell line is used to evaluate the bioactivity of recombinant human bone morphogenetic protein-2 (rhBMP-2).
Storage Conditions liquid nitrogen vapor phase
Derivation

Cells from W++ mouse bone marrow were cloned by limiting dilution to obtain the W-20-17 cell line. RefThies RS. Recombinant human bone morphogenetic protein-2 induces osteoblastic differentiation in W-20-17 stromal cells. Endocrinology 130: 1318-1324, 1992. PubMed: 1311236

Genes Expressed
alkaline phosphatase induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) RefThies RS. Recombinant human bone morphogenetic protein-2 induces osteoblastic differentiation in W-20-17 stromal cells. Endocrinology 130: 1318-1324, 1992. PubMed: 1311236
Comments

Cells used in the W-20-17 assay should not be carried in continuous culture. A working cell bank should be prepared with approximately 5 x 105 cell/mL. Cells should be thawed before each assay.

Recombinant human bone morphogenetic protein-2 (rhBMP-2) increased alkaline phosphatase activity in W-20-17 cells in a dose-responsive manner without an effect on proliferation. RefThies RS. Recombinant human bone morphogenetic protein-2 induces osteoblastic differentiation in W-20-17 stromal cells. Endocrinology 130: 1318-1324, 1992. PubMed: 1311236
Bone morphogenetic protein-3 (BMP-3) was unable to induce an osteogenic response in W-20-17 cells. RefThies RS. Recombinant human bone morphogenetic protein-2 induces osteoblastic differentiation in W-20-17 stromal cells. Endocrinology 130: 1318-1324, 1992. PubMed: 1311236 

Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose, 90%; heat-inactivated 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. Subculture every 3 to 4 days.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-053mM 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 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 gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: At subculture.

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.

Cryopreservation
Culture medium 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Name of Depositor Genetics Institute, Inc.
References

Wang W, et al. Ski represses bone morphogenic protein signaling in Xenopus and mammalian cells. Proc. Natl. Acad. Sci. USA 97: 14394-14399, 2000. PubMed: 11121043

Thies RS. Recombinant human bone morphogenetic protein-2 induces osteoblastic differentiation in W-20-17 stromal cells. Endocrinology 130: 1318-1324, 1992. PubMed: 1311236

Alden TD, et al. In vivo endochondral bone formation using a bone morphogenetic protein 2 adenoviral vector. Hum. Gene Ther. 10: 2245-2253, 1999. PubMed: 10498255

Bahamonde ME, Lyons KM. BMP3: to be or not to be a BMP.. J. Bone Joint Surg. Am. 83-A: S56-S62, 2001. PubMed: 11263666

Standard Test Method for In Vitro Biological Activity of Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) Using the W-20 Mouse Stromal Cell Line. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 2131-02 (Reapproved 2007)e1.

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.

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

Wang W, et al. Ski represses bone morphogenic protein signaling in Xenopus and mammalian cells. Proc. Natl. Acad. Sci. USA 97: 14394-14399, 2000. PubMed: 11121043

Thies RS. Recombinant human bone morphogenetic protein-2 induces osteoblastic differentiation in W-20-17 stromal cells. Endocrinology 130: 1318-1324, 1992. PubMed: 1311236

Alden TD, et al. In vivo endochondral bone formation using a bone morphogenetic protein 2 adenoviral vector. Hum. Gene Ther. 10: 2245-2253, 1999. PubMed: 10498255

Bahamonde ME, Lyons KM. BMP3: to be or not to be a BMP.. J. Bone Joint Surg. Am. 83-A: S56-S62, 2001. PubMed: 11263666

Standard Test Method for In Vitro Biological Activity of Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) Using the W-20 Mouse Stromal Cell Line. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 2131-02 (Reapproved 2007)e1.

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.