Using image-based flow cytometry to monitor satellite cells proliferation and differentiation in vitro

Muscle satellite cells can proliferate and differentiate into myocytes; this biological function has important implications for muscle development, aging, repair, and wasting diseases. Established analytical methods, including western blotting, PCR, and immunohistochemistry, has been used to characterize the stages of satellite cells growth and development but there is currently a lack of methods to simultaneously monitor cell proliferation and identify changes in cell properties (i.e., expression of myogenic regulatory factors) for each individual cell during the progression to become myocytes. Image-based flow cytometry has the capability to simultaneously obtain morphometric data, monitor cell division, and detect expression of multiple proteins of interest. In this article we demonstrate the use of image-based flow cytometry and cell-trace dye to monitor satellite cells proliferation and to identify different stages of satellite cell differentiation.