Enhanced cloning efficiencies of murine hybridomas using human plasma supplemented medium

J Immunol Methods. 1994 May 16;171(2):253-8. doi: 10.1016/0022-1759(94)90045-0.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Division / drug effects
  • Clone Cells / drug effects
  • Culture Media, Conditioned
  • Humans
  • Hybridomas / cytology*
  • Hybridomas / drug effects
  • Hybridomas / immunology
  • Interleukin-6 / blood
  • Interleukin-6 / pharmacology
  • Leukemia, Lymphocytic, Chronic, B-Cell / blood*
  • Mice
  • Mice, Inbred BALB C

Substances

  • Culture Media, Conditioned
  • Interleukin-6