Culturing and Differentiating C2C12 Cells
From Bridges Lab Protocols
Revision as of 18:19, 15 August 2012 by Davebridges (Talk | contribs) (added differentiation protocol)
Growth and Maintenance
- 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.
- Split cells normally 1/10 or 1/20 into 10% FBS with PSG (see Splitting Cells
- Try to split at 70-80% confluence.
- Cells need to be split every other day, if they are not ready refeed them on the second day.
Differentiation
- When cells reach 80-90% confluence switch to media with 2% horse serum.
- Replenish with fresh media every other day.
- Myotube formation is stimulated when the medium is supplemented with 2% horse serum instead of fetal bovine serum.
see http://www.stanford.edu/group/blau/protocols/c2lineprotocol.html and http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=crl-1772&Template=cellBiology for more details.
