A technique for rapid isolation of bone marrow mononuclear cells using Ficoll-Metrizoate and the IBM 2991 blood cell processor

Abstract

Marrow from 7 normal donors and patients was layered onto Ficoll-Metrizoate (FM) under pressure in the IBM 2991 blood cell processor to isolate the mononuclear cell (MNC) population prior to allogeneic transplantation or cryopreservation. This separation method, which takes < 90 min, was a further development since the use of the IBM 2991 to produce a concentrated marrow buffy coat for infusion. By adding FM to the system, marrow stem cells were further concentrated in a small volume with removal of unwanted granulocytes and red blood cells. This facilitated the in vitro treatment of marrow with monoclonal antibodies or drugs, for either the selective elimination of malignant cells prior to autologous bone marrow transplantation (BMT) or T lymphocytes in an attempt to prevent graft vs. host disease (GvHD) in allogeneic BMT. Five of the 7 marrows processed by this procedure have been infused into lethally irradiated recipients with engraftment (allogeneic); the other 2 marrows have been cryopreserved.