title: Measurement of proliferative responses of cultured lymphocytes.
abstract: Measurement of proliferative responses of human lymphocytes is a fundamental technique for the assessment of their biological responses to various stimuli. Most simply, this involves measurement of the number of cells present in a culture before and after the addition of a stimulating agent. This unit contains several different prototype protocols to induce proliferation in lymphocytes following exposure to mitogens, antigens, allogeneic or autologous cells, or soluble factors. Each of these protocols can be used in conjunction with an accompanying protocol, which contains methods to determine cell proliferation by incorporation of [(3)H]thymidine into DNA by nonradioactive methods, e.g., reduction of tetrazolium salts (MTT or WST-1). These protocols provide an estimate of cell proliferation indirectly by measuring DNA synthesis, and cell metabolic activity in an entire cell population, but no data on individual cells is obtained. A protocol for CFSE labeling allows direct detection of single proliferating cells and facilitates the quantification of cell divisions by flow cytometry according to the respective CFSE-dilution, and following costaining with fluorescent labeled antibodies, the characterization of subpopulations in the cell culture.
authors.affiliation: GMBB, National Human Genome Institute, Bethesda, Maryland, USA.
authors.forename: Linda Mesler
authors.initials: LM
authors.lastname: Muul
authors.forename: Guido
authors.initials: G
authors.lastname: Heine
authors.forename: Christopher
authors.initials: C
authors.lastname: Silvin
authors.forename: Stephen P
authors.initials: SP
authors.lastname: James
authors.forename: Fabio
authors.initials: F
authors.lastname: Candotti
authors.forename: Andreas
authors.initials: A
authors.lastname: Radbruch
authors.forename: Margitta
authors.initials: M
authors.lastname: Worm
mesh_terms: D000941:Antigens / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D018655:Lymphocyte Count; D008214:Lymphocytes / Q000166:cytology* / Q000187:drug effects; D008222:Lymphokines / Q000494:pharmacology; D008934:Mitogens / Q000494:pharmacology
chemical_list: D000941:Antigens; D008222:Lymphokines; D008934:Mitogens