The determination of lymphoid cell chimerism using peripheral blood lymphocytes from murine bone marrow chimeras.
A simple, rapid and accurate method was devised for deteriming lymphoid cell chimerism in bone marrow-reconstituted mice. Chimeras were produced by reconstituting lethally irradiated mice with semi-allogeneic bone marrow cells. Lymphocytes from the peripheral blood of individual chimeric mice were purified by sedimentation in dextran solution and differential flotation in Ficoll-Hypaque gradients. From 250--500 microliters of blood, 1--7 X 10(5) cells were routinely obtained. The extent of chimerism was determined serologically by using peripheral blood lymphocytes as target cells in a dye exclusion microcytotoxicity assay. Using this new technique, approximately 80% of the reconstituted mice were found to be repopulated with lymphocytes of the donor type.
D000818:Animals; D001854:Bone Marrow Cells; D003601:Cytotoxicity Tests, Immunologic; D008214:Lymphocytes; D008722:Methods; D051379:Mice; D008832:Microchemistry; D011828:Radiation Chimera