In vitro Differentiation of Embryonic Stem Cells to Muscle like Cells after Treatment with Muscle Extract

One goal of embryonic stem cells (ESCs) research is the development of specialized cells such as neurons, heart muscle cells, endothelial cells of blood vessels, etc. Thus, the directed derivation of ESCs is vital to the ultimate use of such cells in the development of new therapies. This study was conducted to describe the conditions that induce differentiation of inner cell mass (ICM) derived ESCs into muscle like cells. Blastocysts were recovered from female s albino mice of strain Balb c at day 3.5 after natural mating had taken place, cultured in MEM medium supplemented with 20% FCS in the presence of mitotically inactivated feeder layer attached to gelatin layer as a substrate at 37C˚ and 5% CO2. Within two days of the culture, the blastocyst were erupted from the Zona Pellucid and attached by their equatorial pole to the underlying substrate and the ICM, visible as a round dense cell mass in the center of the outgrowth of the trophoblastic cells. Within seven days, the ICM proliferated and increased in their girth, and numbers of ESCs were observed to be librated from the ICM to the surrounding area. When these ESCs were treated with mouse embryonic skeletal muscle extract (ESME) for twelve days, they differentiated and appeared as smooth muscle like cells and expressed positive response against primary monoclonal antibody anti myosin. We can conclude that mouse ESME plays an important role to direct differentiation of ESCs to the muscular pathway. .