Purification of Primary Myoblast by Pre-plating
Material
- Myoblast (containing fibroblast)
See Prepration of primary myoblast by MACS or pre-plating. - Growth medium for myoblast (Ham's/F10, 20% FBS, 2 ng/ml bFGF, PS)
- Fungizone (Amphotericin B) (250 ug/ml, Gibco 15290-018, -20C)
Procedure
- Trypisinize cell.
- Suspend cell in growth medium.
- OPTIONAL: Add 2.5 ug/ml Fungizone (1/100 of stock).
- Seed on non-coated culture dish.
- Incubate at 37C for 60-90 min.
- Check dish whether fibroblast is attaching and myoblast is floating.
- Move supernatant (myoblast-rich) to collagen-coated dish.
- OPTIONAL: Add growth medium to non-coated dish (fibroblast-rich) to keep remaining myoblast.