Multiple stimuli, such as cytokine treatment, can affect cell proliferation. Cell proliferation can occur in response to many stimuli such as cytokine exposure or a variety of other processes.
Cell proliferation dyes
BD Biosciences offers BD Horizon™ Violet Proliferation Dye 450 (VPD450) and BD Horizon™ CFSE for the detection of cell proliferation with the violet laser and blue laser, respectively, which facilitates the use of larger panels. This allows the determination of more data from limited samples using multicolor flow cytometry. Both proliferation dyes are nonfluorescent esterified dyes. The ester group allows the dye to enter the cell. Once the dye is inside the cell, esterases cleave off the ester group to convert the dye into a fluorescent product and trap it inside the cell. With each replication event the amount of dye in the cell is decreased, leading to a characteristic pattern.
BD Horizon™ Cell Proliferation Dyes freely enter a cell. Once inside the cell, the dyes are cleaved by nonspecific esterases and release a fluorescent molecule, which becomes trapped inside the cell.
Concentration of VPD450 and cell-cycle kinetics on mouse spleen stimulated with anti-CD3e and anti-CD28. C57 Black/6 splenocytes were either loaded with varying concentrations of BD Horizon™ VPD450, DMSO, or left as untreated controls, then stimulated with anti-CD3e and anti-CD28 for two days. Cells were pulsed with BrdU prior to harvesting, then stained with APC anti-BrdU and 7-AAD (Cat. No. 552598). The top panels (A–C) illustrate APC antiBrdU and 7-AAD staining. The bottom panels (D–F) illustrate the corresponding VPD450 histograms. The control cells (Cells– ) (Panel A) and the 1-μM VPD450- loaded cell population (Panel C) demonstrated a similar percentage of BrdU+ cells (40.7% and 39.8%, respectively). Higher concentrations of dye can negatively impact cell proliferation (data not shown). To confirm that the DMSO (which is used as a solvent for VPD450) is not responsible for a decrease in proliferation, a DMSO group was included (Panel B). DMSO-treated cells incorporated a similar percentage of BrdU compared to the Cells– group and the 1-μM VPD450-loaded cell populations.
