Removal of rat dendritic cells from single cell suspensions by passage through columns of Sephadex G-10.
Rat T lymphocytes require dendritic cells as accessory cells in order to respond to the mitogen sodium periodate. Passage of a lymph node cell suspension through a column of Sephadex G-10 reduced the mitogenic response by greater than 90%, despite a cell recovery of 75%. An even greater reduction in the response occurred after a second passage over Sephadex G-10; addition of purified dendritic cells restored the response. Lymph node cell suspensions and preparations enriched in dendritic cells were nearly depleted of these cells by passage over Sephadex G-10. After passage of lymph node cells over Sephadex G-10, a limited number of retained cells could be recovered; these included both dendritic cells and macrophages. Enumeration of macrophages in the various passed and retained fractions by non-specific esterase staining of smears confirmed that macrophages were effectively removed from lymph node cell suspensions by repeated passage over Sephadex G-10. Cell preparations that pass through Sephadex G-10 are therefore depleted of both dendritic cells and macrophages.
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002850:Chromatography, Gel / Q000379:methods; D004261:DNA Replication; D003911:Dextrans; D005260:Female; D008198:Lymph Nodes / Q000166:cytology*; D008213:Lymphocyte Activation; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology
D003911:Dextrans; C025614:sephadex