Immunomagnetic purging of bone marrow grafts for autologous transplantation in neuroblastoma

Bone marrow grafts of 5 patients with stage IV neuroblastoma and one patient with stage IV retinoblastoma were harvested during complete remission and in vitro processed for autologous transplantation. Because of the risk to reinfuse metastatic cells, the grafts were immunomagnetically purged. After preparation of the buffy coat fraction, mononuclear cells (mnc) and progenitor cells (CFU-GM) were enriched by density centrifugation and incubated successively with a cocktail of five tumour reacting monoclonal antibodies and magnetic microparticles. Target cells were removed by a set of samarium cobalt magnets. The purged grafts were stored in liquid nitrogen. After the complete procedure, an average of 16% nucleated cells of the initial harvest could be recovered. This corresponds with a mean number of 0.5 X 10(8) mnc and 2.4 X 10(4) CFU-GM per kg body weight being available for grafting. So far, 5 patients received their purged marrow graft and showed a sustained take.