Brugia Malayi - Clark Science Center - Smith College

Two-dimensional Analysis of C2C12 Muscle Cell Line Development
Chelby Wakefield
C2C12 cells are a line of mouse thigh muscle stem cells used to study the development of skeletal muscle in vitro. These cells start
out as myoblasts that are committed to forming skeletal muscle. In low serum containing growth medium these cells exit the cell
cycle and fuse together to form multinucleated early myotubes and eventually late myotubes which are the precursors to mature
muscle fibers. This project focused on the differential proteome profiling of these cells as they progress from myoblasts to early
and late myotubes.
The cells were grown and harvested at the appropriate stages (days 0, 4, and 10). Cellular extracts were then separated in
two dimensions (2D); isoelectric focusing (IEF) and molecular weight. Isoelectric focusing separates proteins according to their
isoelectric point, pI; the pH at which a protein carries no net charge. The second dimension separated proteins according to their
molecular weight by SDS gel electrophoresis. Each spot on the gel represents one or more different proteins (Figure 1). The 2D
gel results yielded changes in protein abundance between the three developmental stages; they were visualized by digital scanning
and analyzed by PDQuest. Spots differing both by two-fold increase or decrease, as well as being statistically significantly different
(p