ABSTRACT
This chapter investigates the use of magnetic monosized particles and anti-T cell monoclonal antibodies for the removal of T cells from human bone marrow cell suspensions. Magnetic monosized M-450 polymer particles coated by physical adsorption with goat antimouse immunoglobulin were effective in removing T lymphocytes from human bone marrow cell suspensions, incubated with the anti-Leu 5 and anti-Leu 2a monoclonal antibodies. Bone marrow was obtained from healthy adult normal donors of both sexes by multiple aspirations from the posterior iliac crest. The separation system was sterilized by washing with 70% alcohol and UV irradiation and washed with phosphate-buffered saline supplemented with 1% fetal calf serum. Eluted cells were characterized by functional and phenotypic tests. Proliferative response to PHA-P and to allogeneic mononuclear leukocytes in mixed lymphocyte culture were absent in the eluted fraction. The separation procedure is carried out in a sterile continuous-flow system, using chambers with permanent samarium-cobalt magnets and a peristaltic pump.
