Abstract

Skeletal muscles in vivo are a highly organized tissues consisting of bundles of myofibers which have preceded by the fusion of myoblasts to form myotubes. Skeletal muscles are of great significance due to their critical involvement in locomotion, metabolism and dynamic activities of the body. Due to these reasons it becomes important to study the biology of skeletal muscle cell and tissues. In vitro tissue models with physiological efficacy provide us with a way to go forward.  One of the key strategies to form skeletal muscle in vitro is ensuring the alignment of skeletal myoblasts by applying passive mechanical stimulus using mechano-topographical cues on culture substrates, facilitating myoblast fusion and differentiation. In the current study we have used OTS (Octadecyltrichlorosilane) /APTES (3 aminopropyltriethoxy silane) micropatterned glass substrate environment to apply topographical constrains on primary skeletal muscle cell progenitors derived from rat hind limb muscles. We performed a comparative study of the overall process of myoblast differentiation in between micropatterned and standard culture plate environments. Standard microscopic techniques like bright field microscopy, fluorescence microscopy and live cell imaging were used for image data. The data obtained was analyzed using Image processing software to infer fusion index and maturation index. The results provided important insights into the fusion, maturation and the overall alignment of the myotubes indicating the superiority of micropatterned substrates over standard culture environment.