Isolation of viable melanoma cells from surgically removed lesions using dishes coated with monoclonal antibody to a high molecular weight melanoma associated antigen

The panning methodology has been applied to isolate viable human melanoma cells from surgically removed lesions. In this procedure a monocellular suspension mechanically prepared from a biopsy is incubated in plastic dishes coated with the monoclonal antibody (MoAb) 225.28S to a membrane bound high molecular weight-melanoma associated antigen (HMW-MAA). The melanoma nature of the cells growing in MoAb 225.28S coated dishes is indicated by the specific reactivity with anti-HMW-MAA MoAb, by its detection in spent culture medium and by morphological criteria. The specificity of the procedure is proven by the lack of growth of cells seeded in fetal calf serum (FCS) coated dishes, as well as of cells lacking the HMW-MAA in MoAb 225.28S coated dishes. The adherence of melanoma cells to plastic dishes is influenced by the concentration of the plastic bound MoAb 225.28S and by the incubation time. Melanoma cells isolated by adherence to MoAb 225.28S coated plates do not display any detectable change in their growth curve and colony forming ability. The ready availability of melanoma cells isolated from surgically removed lesions will greatly facilitate the characterization of the interaction between host's immune system and tumor cells and the screening of anti-tumor agents in preclinical tests.