A procedure for the isolation of highly purified populations of B cells, T cells and monocytes from human peripheral and umbilical cord blood.
A sequential separation protocol is described which reproducibly yields biologically active populations of B cells, T cells and monocytes from human peripheral and umbilical cord blood. The purification protocol employed T lymphocyte rosetting with unmodified sheep red blood cells, monocyte adherence to microexudate-coated flasks and anti-immunoglobulin panning of B cells. Population purity was determined by cytofluorographic light scatter analysis, expression of cell surface markers, esterase activity and mitogen responsiveness. The sequential protocol reproducibly yielded viable and functionally active cell populations of greater than 95% purity from a single starting population of mononuclear cells.
D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes; D002469:Cell Separation / Q000379:methods; D004950:Esterases / Q000097:blood; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D008027:Light; D008213:Lymphocyte Activation; D009000:Monocytes; D012542:Scattering, Radiation; D013601:T-Lymphocytes
D000954:Antigens, Surface; D004950:Esterases