I have been having trouble consistently growing and differentiating primary human skeletal muscle cells in the seahorse 96 well plates. The cells will grow to confluency and differentiate into myotubes with the addition of differentiation media, but after a few days they clump up and pull off the plate. This ends up leading to really sloppy inconsistent data. Has anyone ever even worked with primary human skeletal muscle cells in seahorse plates before? I have looked through lots of literature but found no papers using primaries. Just a lot of papers using C2C12s and other cell lines.