Harvest and Isolate Skeletal Muscle Satellite Cells from Mice
Skeletal muscle tissue exhibits many plastic properties, changing its size or function in response to a variety of stressors. This adaptability is due, in part, to satellite cells, a type of multipotent adult stem cell that are found between the sarcolemma and basement membrane of muscle fibers. These cells, while normally quiescent, are activated by mechanical strain or injury. Whe activated, they are responsible for creating new muscle fibers and/or combining with existing myofibers to facilitate growth and repair. After an injury, satellite cells begin to proliferate and differentiate into myoblasts to facilitate the repair and regeneration of muscle tissue. Further understanding the contributions of satellite cells to the plasticity of skeletal muscle may lead to important medical contributions for a number of diseases associated with the loss of skeletal muscle mass or function, such as muscular dystrophy. This study sought to accomplish two specific aims, the first of which was to harvest and isolate satellite cells from intact murine muscle, and subsequently grow these cells into viable myotubes in culture. The second aim of these studies was to characterize the cultured myotubes and compare aspects of their anabolic profiles to tissue from the same animal at the time of tissue harvest. Outcomes for these studies include analyzing signaling outcomes between cultured and intact muscle in an effort to understand morphological/metabolic differences between these conditions. The central hypothesis for this study is that phenotypic expression of skeletal muscle (fiber type, metabolic profiles, anabolic profiles) will persist in cell culture. If the harvested cells maintain similar characteristics to the intact tissue, these methodologies can be used to better understand mechanisms of muscle disease states in cell culture, and potentially, translate our findings to intact living organisms. Ultimately, if successful, these methodologies could be adapted to isolating satellite cells from biopsied human skeletal muscle tissue, and subsequently invested differentiated myotubes that retain specific muscle abnormalities of the donor. This would allow for the ability to harvest and maintain human cells expressing a desired trait, without the constant need to obtain muscle biopsies from new subjects.
Nguyen, Branden (2017). Harvest and Isolate Skeletal Muscle Satellite Cells from Mice. Undergraduate Research Scholars Program. Available electronically from