In order to establish a protocol for stem cell differentiation, we experiment on different methods in C2C12 cells. To begin, C2C12 cells transduced with the myoD promoter were grown to full confluency. When differentiation began, the control cells grown horizontally were compared to the experimental cells grown vertically. The cells grown vertically were generally much longer than those grown horizontally. One of the major conditions of differentiation is when the promoters are activated. If the promoters activate faster for cells grown vertically, this could explain the longer multinucleated cells. The promoter gene, MyoD, was tested for flourescense indicating activation to determine inconsistencies in timing between the control and experimental cells. After imaging each plate containing an individual promoter gene on days 0, 3, 5, and 7 of differentiation, the observed fluorescence began on the control plate first. It is possible that the later activation of the promoter genes allows the cells to grow longer prior to differentiation explaining why cells grown on the experimental plate are longer.
