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C2C12
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Biosafety Level:
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1
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frozen
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Medium & Serum:
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See Propagation
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adherent
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Mus musculus (mouse)
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myoblast
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Strain: C3H
Tissue: muscle
Cell Type: myoblast;
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In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
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transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
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ATCC 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%.
Temperature: 37.0°C
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Protocol: 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.
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Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
Inoculate at a cell concentration between 1.5 X 10 exp5 and 1.0 X 10 exp6 viable cells/75 cm2.
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Incubate cultures at 37°C.
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