A RAPID METHOD FOR SEPARATING TUMOR INFILTRATING CELLS AND TUMOR-CELLS FROM COLORECTAL TUMORS

As solid tumours are composed of a heterogeneous mixture of cells the role of any particular cell type is difficult to analyse. A rapid method of sorting freshly disaggregated highly viable single cell suspensions of colorectal tumours has therefore been developed. Infiltrating cells were initially sorted from tumour cell suspensions using magnetic beads coated with a cocktail of monoclonal antibodies recognising leucocyte common antigen, CD45 (lymphocytes), CDw14 (macrophages and granulocytes) and Thy-1 antigen (stromal cells). Between 4-10 x 10(6) cells, 70-87% pure, were rapidly sorted. The tumour cells were then sorted from the remaining cell suspension using a cocktail of monoclonal antibodies which recognise 100% of colorectal tumours. Between 2-7 x 10(6) tumour cells, 70-87% pure, were rapidly sorted. Both populations of cells were 95% viable and able to grow in vitro.