STEMdiff Myogenic Progenitor Kit

This serum-free supplement system from STEMCELL Technologies is designed to differentiate human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) into myogenic progenitor cells. The resulting cells are characterized by myogenic markers including CD56 and CD82. The kit is compatible with hESCs and hiPSCs maintained in mTeSR™1, mTeSR™ Plus, or TeSR™-E8™.

The differentiation protocol spans 30 days. On Day -1, hPSCs are plated on Corning® Matrigel®-coated plates at a density of 30,000 - 40,000 cells/cm². Daily full-medium changes use Supplement A on Days 0 - 2, Supplement B on Days 2 - 4, Supplement C on Days 4 - 6, and Supplement D from Day 6 through Day 29. On Day 30, the culture contains PAX7+ MyoD+ myogenic progenitors that can be sub-cultured for downstream use.

These progenitor cells can be culture-expanded for 5+ passages using the MyoCult™-SF Expansion Supplement Kit (Human) and further differentiated into functional, multinucleated MyHC+ myotubes using the MyoCult™ Differentiation Kit (Human). hPSC-derived myotubes generated through this workflow display organized sarcomeric structures, as indicated by alpha-actinin staining, and exhibit spontaneous contractile activity as recorded by microelectrode assay. Multiple hPSC lines differentiated with this kit showed high fusion indices similar to primary human myoblasts.