Comparison of fluorescence intensity of Hoechst 33342 – stained EMT6 tumour cells and tumour-infiltrating host cells

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Abstract
Hoechst 33342 is a fluorescent dye used for cell selection from tumours based upon intratumour location. When the dye is administered i.v. to tumour-bearing animals, cellular fluorescence is directly related to the proximity of cells to blood vessels. The present study compared inherent Hoechst fluorescence between in vitro-stained EMT6/Ro (mouse mammary sarcoma) cells and host cells, to determine if these populations have different staining characteristics that may influence cell selection procedures. Tumour cell fluorescence exceeded host cell staining by 8-fold when pure cell populations (EMT6/Ro monolayer cells, mouse spleen and peritoneal cells) were compared, and 3-fold for tumour cell-enriched and host cell-enriched populations from solid tumours. Inherent uptake of HO 33342 appeared to be correlated with cell volume. These differences in inherent dye uptake between host and tumour cells were found to be minor in comparison to the fluorescence gradient between the 10% brightest and 10% dimmest (78-fold) cell populations from in vivo-stained tumours.