2.6. Immunofluorescence Staining of Differentiated Myoblasts

The differentiation of mouse myoblast cells within both hydrogels was studied in a glass confocal dish (12mm) by immunofluorescence analysis. C2C12 (30,000 cells/plate) were embedded in different hydrogels. After 8 days of differentiation, 4% paraformaldehyde solution was used for cells fixation. After 20 min incubation at room temperature, the cells were permeabilized and labeled with primary anti-MHC (1:300 PBS) for 1 h followed by 1 h incubation with secondary anti-mouse IgG-fluorescein isothiocyanate and DAPI. The myotube formation was observed with fluorescence confocal microscopy (Zeiss LSM 710 Inverted Confocal Microscope, Germany).