The epitome that cell carrier serves solely as passive vehicles has become outdated. It is now evident that the carrier microenvironment also contributes in the regeneration process. In this study, a combination of alginate, pluronic F127 and extracellular matrix (ECM) component, hyaluronic acid (HA) based scaffold has been prepared for in situ gelling vehicles for muscle cells. ECM incorporated blended hydrogel showed enhanced uniform distribution of muscle cells in a nude mouse model forming the scaffold in situ allowed the muscle cells to proliferate efficiently, indicating that a pluronic F127/alginate/HA matrix provided a beneficial environment for cellular growth and expansion. The formation of gel beneath the skin of nude mice was confirmed using optical coherence tomography (OCT). OCT has been used to visualize the in situ localization of cells as well. This in situ gelation is found to be advantageous for regenerative applications due to the absence of toxic solvents or co-polymerization agents; besides the handling process is simple. This study demonstrates that an in situ blended hydrogels enables the favorable settlement of cells and satisfactory cell delivery for muscle regeneration applications.