Abstract
Mouse peritoneal exudate cells induced by casein enhanced in vitro antibody production rate per cell of a hybridoma in co-culture. Culture supernatant of the exudate cells also enhanced three-fold the antibody productivity when added to cultures of a hybridoma at 10% (v/v). Hence the enhancement of antibody productivity by the exudate cells seemed to be caused by soluble enhancing factors secreted by the exudate cells. The exudate cells maximally secreted the enhancing factors when harvested from mice on day 4 of the induction period following the injection of casein. A semi-continuous culture of the hybridoma demonstrated the applicability of the culture supernatant to enhance antibody production by producing a two-fold increase over the control for seven days when supplemented with the supernatant at 5%. Significant amounts of interleukin-6 were detected in culture supernatant of the exudate cells. Interleukin-6 obtained from other sources enhanced the antibody productivity two-fold when added to the hybridoma culture at the concentration of 5 unit/ml. Interleukin-6, therefore, is expected to be one of the principal antibody enhancing factors secreted by the exudate cells. Other interleukins examined, that is, interleukin-1 to-5 did not enhance the antibody productivity.
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AL-RubeaiM, MillsD and EmeryAN (1990) Electron microscopy of hybridoma cells with special regard to monoclonal antibody production. Cytotechnology 4: 13–28.
ConlonPJ, HenneyCS and GillisS (1982) Cytokine dependent thymocyte responses characterization of interleukin 1 and interleukin 2 target sub populations and mechanism of action. J. Immunol. 128: 797–801.
GodingJW (1980) Antibody production by hybridomas. J. Immunol. Methods. 39: 285–308.
HiranoT, YasukawaK, HaradaH, TagaT, WatanabeY, MatsudaT, KashiwamuraS, NakajimaK, KoyamaK, IwamatsuA, TsunasawaS, SakiyamaF, MatsuiH, TakaharaY, TaniguchiT and KishimotoT (1986) Complementary DNA for a novel human interleukin (BSF-2) that induces B lymphocytes to produce immunoglobulin. Nature 324: 73–76.
KishimotoT, TadaT and YamamuraY (1987) Immunology 87, Nakayama shoten, Tokyo.
MertenO-W, KellerH, CabanieL, LenoM and HardefeltM (1990) Batch production and secretion kinetics of hybridomas: Pulse-chase experiments. Cytotechnology 4: 77–89.
MillerWM, BlanchHW and WilkeCR (1988) A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: effect of nutrient concentration, dilution rate, and pH. Biotechnol. Bioeng. 32: 947–963.
NakamuraS, GotoF, GotoK and YoshinagaM (1982) Physicochemical characterization of a PMN-derived soluble factor that enhances lymphocyte DNA synthesis. J. Immunol. 128: 2614–2621.
ReuvenyS, VelezD, RiskeF, MacmillanJD and MillerL (1985) Production of monoclonal antibodies in culture. Dev. Biol. Stand. 60: 185–198.
SuzukiE and OllisDF (1990) Enhanced antibody production at slowed growth rates: Experimental demonstration and a simple structured model. Biotechnol. Prog. 6: 231–236.
YamadaK, IkedaI, SugaharaT, ShirahataS and MurakamiH (1990) Enhancement of immunoglobulin production of human-human hybridomas by immunoglobulin production stimulating factors. In: MurakamiH (ed.) Trends in Animal Cell Culture Technology (pp. 291–297). Kodansha, Tokyo.
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Mikami, T., Makishima, F. & Suzuki, E. Enhancing effect of mouse peritoneal exudate cells and their products on antibody productivity of hybridoma cells: Application of in vivo factors to in vitro culture. Cytotechnology 7, 93–101 (1991). https://doi.org/10.1007/BF00350915
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DOI: https://doi.org/10.1007/BF00350915