Collagenase digestion and mechanical disaggregation as a method to extractand immunophenotype tumour lymphocytes in cutaneous T-cell lymphomas

Various enzymatic or mechanical methods have been proposed in the past to d issociate cells from different solid tissues. An automated mechanical disag gregation device (Medimachine(TM)) has recently been proposed. Unfortunatel y, most of these techniques are associated with a high cellular damage and a low cell recovery and are difficult to apply to skin biopsies. In this pa per, we propose a combined enzymatic and mechanical method based on Medimac hine(TM), useful for the isolation of skin infiltrating T-lymphocytes from small cutaneous biopsies. As this method is easy and allows for a more corr ect qualitative and quantitative cytofluorimetric analysis of the lymphocyt e subsets, it may be useful in the immunophenotyping of cutaneous T-cell ly mphomas.