title: Enhanced cloning efficiencies of murine hybridomas using human plasma supplemented medium.
abstract: Normal human plasma and plasma from patients with chronic lymphocytic leukemia (CLL) were used as growth supplements for the cloning of murine hybridomas. Basal medium, consisting of DMEM with 5% foetal calf serum (FCS) was conditioned with supernatant from a known human IL-6-secreting cell line, BRI-6 (BRI-6-CM), normal human plasma (NHP-DMEM), and plasma from patients with CLL (CLL-DMEM). When compared to conventional feeder layers of macrophages, thymocytes, splenocytes and to feeder CLL cells and BRI-6-CM the numbers of clones formed by growing hybridomas in CLL-DMEM was greatly enhanced with a corresponding increase in the number of antibody-producing clones, as determined by ELISA. NHP-DMEM also enhanced the cloning efficiency. All CM and plasma supplemented medium were examined for the presence of interleukin-6 (IL-6). Of eight CLL plasma samples examined only two had elevated IL-6 levels.
authors.affiliation: National Cell and Tissue Culture Centre/BioResearch Ireland, School of Biological Sciences, Dublin City University.
authors.forename: N
authors.initials: N
authors.lastname: Quinlan
authors.forename: R
authors.initials: R
authors.lastname: O'Kennedy
mesh_terms: D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells / Q000187:drug effects; D017077:Culture Media, Conditioned; D006801:Humans; D006825:Hybridomas / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D015850:Interleukin-6 / Q000097:blood / Q000494:pharmacology; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000097:blood*; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D017077:Culture Media, Conditioned; D015850:Interleukin-6