Maturation Profiling of Human Monocyte-Derived Dendritic Cells (Mo-DCs)

Begin collecting a new sample of the cells of interest for antibody staining by washing the cells at room temperature for five minutes at 300 G with normal break. Distribute the cells into micro tubes. To perform the staining using the desired antibodies, incubate the fluorescence conjugated antibodies in the dark for 15 minutes at room temperature.

Wash the cells with one milliliter of PBS before centrifuging for five minutes at 300 G at room temperature. Add up to 100 microliters of PBS to all the micro tubes. Then resuspend the cells in 300 microliters of 2%paraformaldehyde, and store the tubes in the dark at four degrees Celsius until flow cytometry.

A significant increase was observed in the expression of the antigen presenting molecules MHC-I and MHC-II, and the expression of the CD80 and CD86 co-stimulatory molecules due to sialidase treatment.