Can the proliferative ability of chicken cardiomyocytes be assessed using flow cytometry?

Abstract: The study of the formation of new cardiac muscle cells during postnatal development is a relatively new field. During fetal development, new cells are formed as the heart grows. However, the proliferative ability of postnatal cardiomyocytes is still debated. While several studies have been made on mammals, less is known about the chicken cardiac cells and their postnatal proliferation. As almost all previous studies have used microscopy-based cell counting methods, there has been some limitations on accuracy and amounts of cells that could be counted. The aim of this study is to develop a method for using flow cytometry to analyze proliferative ability of chicken cardiomyocytes and to investigate if any postnatal proliferation exists. For this study, 4 weeks old Red Junglefowl (Gallus gallus) chickens were used for isolating cardiomyocytes. In addition, 19 days old Red Junglefowl embryos were used to asses if a longer incubation time would yield a higher number of proliferative cells. Cells were stained using a commercial EdU imaging kit and analyzed using flow cytometry and imaging flow cytometry. The produced results could not be used for determining the proliferative ability of the cardiomyocytes, but provides crucial information for possible method improvements. In conclusion, this study has laid important groundwork for future studies on the proliferative ability of chicken cardiomyocytes.