A method for enrichment tumor cells from bone marrow was developed to provide even greater sensitivity in detecting tumor cells that have metastasized to marrow. Mixtures of normal marrow and cultured neuroblastoma cells were reacted with anti-cell su...
A method for enrichment tumor cells from bone marrow was developed to provide even greater sensitivity in detecting tumor cells that have metastasized to marrow. Mixtures of normal marrow and cultured neuroblastoma cells were reacted with anti-cell surface monoclonal antibodies, and then magnetic immunobeads coated with secondary antibodies that react with primary antibodies; the bead-target cell conjugates were removed from the mixture with a magnet. Negative selection with a combination of four anti-hematopoietic antibodies (anti-Ia, anti-myeloid, anti-)32 microglobulin, anti-HLA) produced enrichment of tumor cells from marrow (e.g. 1% can be enriched to 9.6%). Equilibrium sedimentation over Ficoll-Hypaque, which also enriched tumor cells about 3.6 fold, followed by removal of most hematopoietic cells using negative selection can concentrate tumor cells one and half logs. Combined with nnmunohistology using anti-neuron specific enolase antibody, It is suggested that this may increase sensitivity to one neuroblastoma cell per 10` normal cells.