During our QC analysis we test every lot of CRL-1772 for myotube formation by allowing the cells to become confluent. The procedure that we use during this testing is: A T75 flask is seeded and should be 80% to 90% confluent on day 3. The cells are then monitored and fed with fresh complete culture medium [500 mL ATCC DMEM (30-2002) supplemented only with 56 mL ATCC FBS (30-2020)] only as needed to keep the cells alive. After a total of 14 days incubation there should be good myotube formation. The myotubes appears as thick tubular structures, sometimes multinucleated.
These cells will differentiate at confluence alone however changing to horse serum after reaching 100% confluency encourages faster myogenesis by reducing the number of growth factors available to the cells. We are aware that myotube formation is enhanced when the medium is supplemented with horse serum instead of FBS, but in our labs we simply use the complete culture medium with no problems.
The differentiation potential is dependent on how the cells have been cultured and subcultured. To prevent the loss of myoblastic cells during regular passaging it is critical that the cells are not allowed to become confluent. The myoblast population of the CRL-1772 cell line will become depleted rapidly if the cultures are allowed to become confluent; this can significantly delay the differentiation of these cells.
|Date Created||12/18/2017 10:35 AM
|Date Updated||12/18/2017 10:36 AM