The physical and mental impact that head and neck skeletal muscle loss has on patients can be significant. So far, the greatest challenge in this area has been the tissue engineering of functional skeletal muscle. Quite often the affect of head and neck skeletal muscle loss is rectified by means of artificial materials and autogenous implants. Nevertheless, this approach has not proven to be too practical. Objectives: to describe an approach to culture skeletal muscle that has a 3-D organization similar to an intact skeletal muscle and to characterize this tissue by specific DNA markers. Methods: satellite cells were isolated from the hind limbs of 2-4 day old rats by protease digestion, and then separated from fibroblasts by differential adhesion. After explantation by culturing in high serum media (DMEM-25% FBS), the isolated satellite cells were transferred into new culture vessels coated with aligned collagen and then cultured in low serum media (DMEM-10% FBS). This media is appropriate for differentiation of satellite cells into myotubes with a parallel arrangement. After 5-7 days the total RNA was extracted from this cultures. In order to characterize this tissue by RT-PCR we made DNA probes (to measure the specific mRNAs) for the muscle specific markers such as, Myogenin, Desmin, Integrin alpha-7 and muscle specific Actin. As a negative control we used mouse liver mRNA for which Desmin was used as a probe. Results: all specific markers show strong bands on agarose gel when compared to the control. Conclusions: we have successfully isolated skeletal satellite cells; and most importantly, we achieved growing those cells in an aligned pattern. Also, with the aid of muscle specific markers, we characterized that those cells are compatible with intact skeletal muscle.

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