Fluorescence-based method is more accurate than counting-based methods for plotting growth curves of adherent cells

摘要

Analysis of CFSE signal decay upon cell proliferation. CFSE passively diffuses into the cells and covalently binds to free amine residues. As cells divide, each daughter cell holds half of the CFSE content present in the mother cell. HEK 293 cells were stained with CFSE and cultured for up to 7 days. Cells were harvested and fixed at different time points and the CFSE fluorescence signal was measured. Consecutive cell divisions lead to progressive CFSE signal decay in the cell population. The experiment was carried out with three technical replicates. CFSE MFI as function of time reveals the exponential CFSE signal decay. The inverse of CFSE MFI values (MFI ) plotted as function of time renders the curve into a sigmoid, similar to the conventional growth curves