Assessment of Cell Migration in vivo Using the Flurorescent Tracking Dye PKH26 and Flow Cytometry

Abstract

The ability to determine the migration of specific cell subpopulations to certain tissue target sites is important in identifying the role of cell mediated immunity in the pathogenesis of autoimmune disorders. The highly fluorescent PKH tracking dyes produce a stable cell membrane labelling and have been used in combination with flow cytometry and multicolour immunofluorescence to determine the involvement of specific subsets of cells in adoptive transfer of insulin-dependent diabetes melitus (IDDM) in the non-obese diabetic (NOD) mouse model (see Beavis and Pennline, J. Immunol. Methods, 170, pp 57–65, 1994). Here, the PKH26 tracking dye (Sigma, PKH26-GL) was used to label donor cells and then to follow their migration to various sites in a recipient mouse for up to 28 days following transfer. This was combined with phenotypic analysis using FITC and APC-labelled antibodies.