Objective: To investigate the myogenic differentiation of laryngeal mucosal mesenchymal stem cells (LM-MSCs) and the possibility of LM-MSCs as new alternative seed cells for laryngeal tissue engineering.
Methods: LM-MSCs were separated from normal epiglottis mucosa and the cell surface markers including CD44, CD105, CD90, CD29, CD34 and CD45 were analyzed through flow cytometry. The osteogenesis and adipogenesis differentiation of LM-MSCs were investigated by oil red staining and alizarin red S staining. Immunofluorescence staining and RT-PCR were used to detect the expressions of myogenic differentiation markers including Myod1, Myogenin and myosin heavy chain (MyHc).
Results: The separated LM-MSCs were in a fibrocyte-like form with long fusiform shape and grew adherent. The expression rates of cell surface markers LM-MSCs were CD44 (100.0%), CD105 (90.4%), CD90(99.9%), CD29 (93.0%), CD34 (0.4%) and CD45(1.3%) respectively. A number of beaded lipid drops and mineral deposition were observed after 14 days of adipogenesis differentiation and 21 days of osteogenesis differentiation. Myod1, Myogenin and MyHc genes appeared after 1 week and 3 weeks of myogenesis differentiation respectively.
Conclusions: The LM-MSCs have the properties of mesenchymal stem cells and could be differentiated into myoblasts, providing with the possibility to repair the damaged vocal cords with LM-MSCs through tissue engineering techniques.