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C2C12; Muscle Myoblast; Mouse (Mus musculus) Item ID: ATCCRL1772

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Organism Mus musculus, mouse
Tissue muscle
Cell Type myoblast
Product Format frozen
Morphology myoblast
Culture Properties adherent
Biosafety Level 1

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.
Strain C3H
Applications transfection host
Storage Conditions liquid nitrogen vapor phase
Images
Derivation This is a subclone (produced by H. Blau, et al) of the mouse myoblast cell line established by D. Yaffe and O. Saxel.
Comments The C2C12 cell line differentiates rapidly, forming contractile myotubes and producing characteristic muscle proteins. Treatment with bone morphogenic protein 2 (BMP-2) cause a shift in the differentiation pathway from myoblastic to osteoblastic. Tested and found negative for ectromelia virus (mousepox).
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Seeding Density It is important that these cells be seeded at a density of only 5.0 X103 viable cells/cm2; refer to the COA for the batch specific information.
Subculturing IMPORTANT - DO NOT ALLOW CULTURES TO BECOME CONFLUENT. Cultures must not be allowed to become confluent as this will deplete the myoblastic population in the culture. Myotube formation is enhanced when the medium is supplemented with 10% horse serum instead of fetal bovine serum.
  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 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).
    4. 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. Inoculate at a cell concentration between 1.5 X 10 5 and 1.0 X 10 6 viable cells/75 cm2. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  6. Incubate cultures at 37°C.
Medium Renewal: Every two to three days
Cryopreservation Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions Temperature: 37.0°C
Name of Depositor B Paterson
References

Qing Y, et al. Inhibitory effects of iron on bone morphogenetic protein 2-induced osteoblastogenesis. J. Bone Miner. Res. 26(6): 1188-1196, 2011. PubMed: 21308772

Chow YH, et al. Improvement of hepatitis B virus DNA vaccines by plasmids coexpressing hepatitis B surface antigen and interleukin-2. J. Virol. 71: 169-178, 1997. PubMed: 8985336

Hsu DK, et al. Identification of a murine TEF-1-related gene expressed after mitogenic stimulation of quiescent fibroblasts and during myogenic differentiation. J. Biol. Chem. 271: 13786-13795, 1996. PubMed: 8662936

Kessler PD, et al. Gene delivery to skeletal muscle results in sustanined expression and systemic delivery of a therapeutic protein. Proc. Natl. Acad. Sci. USA 93: 14082-14087, 1996. PubMed: 8943064

Katagiri T, et al. Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage [published erratum appears in J Cell Biol 1995 Feb;128(4):following 713]. J. Cell Biol. 127: 1755-1766, 1994. PubMed: 7798324

Blau HM, et al. Plasticity of the differentiated state. Science 230: 758-766, 1985. PubMed: 2414846

Yaffe D, Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature 270: 725-727, 1977. PubMed: 563524

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