1988 Volume 11 Issue 4 Pages 346-356
Raji cells derived from Burkitt lymphoma, were mutated in vitro with ethylmethanesulfonate (EMS) and selected in the presence of 1×10-4M of 6-thioguanine (6-TG). The most rapidly growing 6-TG-resistant clone was obtained and named Raji 6-TGR cells. Furthermore, plasmid vector pSV2 neo gene was transfected to Raji 6-TGR cells by liposomes-mediated gene transfer technique. The transfected cells were selected by neo selective medium (10% FCS-RPMI 1640 medium supplemented with 1mg/ml Geneticin). The fastest growing neo-resistant clone, Raji 6-TGRneoR cells were established. The characteristics of Raji 6-TGR-neoR cells were as follows;
1. Raji 6-TGR-neoR cells were deficient in hypoxanthine-guanine phosphoribosyl transferase (HGPRT) activity, because this cell line was sensitive to HAT (H: 1×10-4M, A: 2×10-8_??_1×10-7M, T: 1.6×10-5M) selective medium.
2. Raji 6-TGR-neoR cells did not secrete human Ig in culture medium, when tested by enzyme-linked immunosorbent assay (ELISA).
3. Raji 6-TGR-neoR cells were fused with B cells, PWM stimulated B cells (PWM-B cells) or EBV transformed B cell line (CESS cells), and selected by a new selective medium (HAT-neo). The percentage of cell growth was as follows, B cells (2%), PWM-B cells (5%) and CESS cells (11%).
These results suggest that Raji 6-TGR-neoR cells are suitable for production of human B cell hybridomas in the new selective medium (HAT-neo).
