Skeletal muscle fibers develop from mono-nucleated myoblasts that fuse to form multinucleated myotubes. In embryonic growth, this process occurs concurrently with the formation of the early cartilaginous skeleton and innervation by migrating nerve cells. The goal of my research was to explore co-culture conditions that encourage proliferation, differentiation and maturation of myoblasts to myotubes. A variety of co-culture experiments tested the influence of three basic tissues types (murine neural, cartilage and kidney primary cells) on the formation of myotubes in the C2C12 myoblast cell line. Three plating strategies were used: 1) C2C12 myoblasts were plated first, grown for two days before the addition of a second cell type; 2) both cell types were mixed and plated simultaneously; and 3) C2C12 myoblasts were added to a pre-established, 10 day old neural, cartilage or kidney cell culture. In addition, a parallel set of experiments were treated with all-trans retinoic acid, a potent myogenic activator and embryonic patterning signaling molecule. Myotube formation was consistently highest in C2C12 and cartilage co-cultures across all three plating strategies with a 277% increase in myotube area compared to controls. These effects were further enhanced when grown in 1 µg/mL all-trans retinoic acid. Co-cultures with neural or kidney cells consistently exhibited fewer myotubes when compared to C2C12 controls. It is postulated that the enhanced muscle growth in cartilage co-cultures was due to a chondrocyte-secreted extracellular matrix that facilitated myotube attachment to the substratum. / Graduate
| Identifer | oai:union.ndltd.org:uvic.ca/oai:dspace.library.uvic.ca:1828/3776 |
| Date | 23 December 2011 |
| Creators | Grey, Matthew |
| Contributors | Nahirney, Patrick C. |
| Source Sets | University of Victoria |
| Language | English, English |
| Detected Language | English |
| Type | Thesis |
| Rights | Available to the World Wide Web |
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