A simple method to obtain low density marrow cells for human marrow transplantation.
Removal of more than 99% of the erythrocytes and 74% of the nucleated cells from marrow grafts was achieved by density floatation separation in Percoll gradients with a density of 1.070 g/ml in eight 250-ml tubes, containing up to 3 X 10(9) nucleated cells per gradient. More than 90% of the myeloid and erythroid progenitor cells were recovered in the low density fraction. It appeared mandatory to use a centrifuge with the possibility of a gradual acceleration and deceleration. Twenty-five patients received a marrow graft from a histocompatible sibling after additional lymphocyte depletion by counterflow centrifugation, and 5 patients with T lymphoblastic malignancies received an autograft after in vitro purging with immunotoxins. All evaluable patients engrafted within normal limits, except 1 patient with an autoimmune pancytopenia who responded to steroids and 1 patient with a CMV infection. Four patients died too early for complete evaluation. The described separation method is easy, cheap and requires only 2 h for the complete processing of a marrow graft.
de Witte
T
T
Department of Internal Medicine, University Hospital Nijmegen, The Netherlands.
Plas
A
A
Vet
J
J
Koekman
E
E
Preyers
F
F
Wessels
J
J
D016026:Bone Marrow Transplantation; D002499:Centrifugation, Density Gradient / Q000379:methods; D006801:Humans; D007945:Leukemia, Lymphoid / Q000628:therapy; D011205:Povidone; D013048:Specimen Handling
D011205:Povidone