Motor Endplate Formation from AMDCs

Cultured AMDCs were allowed to reach confluence. Cells were then cultured with differentiation media containing DMEM supplemented with 2% horse serum and 1% PSF-1 for 5 days with media changes every 48 hours. Thereafter, induction media containing agrin (10 nM, R&D systems), neuregulin (2 nM, R&D Systems), and acetylcholine (10 nM, R&D Systems, Minneapolis, MN) was added to induce motor endplate formation. After 5 days in induction culture media, the cells were used for the study. MEE differentiation was confirmed by immunostaining with Alexa Fluor 594 conjugated bungarotoxin (Molecular Probes, Eugene, Oregon) [6 (link)].