Abstract
Chemical agents (DTT, EDTA) and enzymes (collagenase, DNAse) are often used for the generation of a single cell suspension from tissue samples. In this study, flow cytometry was used to examine the effect of chemical agents and enzymes on the expression of cell membrane markers of T lymphocytes from tonsils and peripheral blood. Expression of CD4, CD8, CD25, CD38, L-selectin, CD44, αEβ7 and α4β7 were studied. Incubation of lymphocytes with DTT and EDTA resulted in a decrease of CD38, αEβ7 and α4β7 expression. Incubation with collagenase A and DNAse resulted in a decrease of CD25, L-selectin, αEβ7 and α4β7. The results of this study indicate that a careful interpretation is necessary for phenotypic descriptions of lymphocyte populations obtained by enzymatic isolation techniques.
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