Yamamoto I, Miyai E, Chiba T
Department of Immunochemistry, Faculty of Pharmaceutical Sciences, Okayama University, Japan.
J Immunol Methods. 1993 Feb 26;159(1-2):153-60. doi: 10.1016/0022-1759(93)90153-x.
In order to induce an optimal plaque-forming cell response to sheep red blood cells (anti-SRBC PFC response) in vitro, murine splenocytes have been cultured in RPMI 1640 medium containing 10% fetal calf serum (FCS). In this paper, we report that 10% intact FCS in such a conventional medium can be replaced by 2% modified FCS which has been exposed to insoluble polymer beta-cyclodextrin (CD). The most efficient procedure of treatment of FCS was an incubation of 1/50 diluted FCS with 20 mg/ml polymer beta-CD for 3.5 h at 20 degrees C. The PFC response supported by 2% polymer beta-CD-treated FCS, was the same as that obtained when mixing 10% FCS, and was completely dependent on the existence of antigen (SRBC) and enhanced by 2-mercaptoethanol. The time-course profiles of the PFC responses induced in both cultures were almost the same. The 2% polymer beta-CD-treated FCS-containing medium was also effective in inducing both a primary PFC response to the T cell-independent antigen (trinitrophenyl-lipopolysaccharide) and a polyclonal PFC response stimulated by lipopolysaccharide. Inactive FCS lots (so called deficient lots) also became fully supportive at 2% after treatment with polymer beta-CD. These results indicate that polymer beta-CD-treated FCS is not economical for supporting antibody production, but also useful for those experiments on isolation and analysis of factors derived from cultured lymphocytes.
