u/LavishnessSmooth1608

Very recently, I have attempted to differentiate C2C12 myoblasts into myotubes by waiting until confluency to change the proliferation media to differentiation media. However, around 3 to 4 days after seeding, cells started to form micromasses or cell aggregates before switching to low serum media, as shown in the first two images. Cells are grown on tissue culture-treated plates.
In other plates, the cells also delaminated or detached from the plate, though I suspect this is due to the plates not being treated with Matrigel or poly-l-lysine.
My main concern is the micromass formation. Any recommendations on how to avoid the micromasses and establish a more reproducible culture of C2C12 myotubes are greatly appreciated.