Bovine colostrum ultrafiltrate: an effective supplement for the culture of mouse-mouse hybridoma cells

Abstract

An ultrafiltrate fraction (UF) of bovine colostrum has been successfully used as a cell culture supplement for growth and monoclonal IgG antibody production of cultured mouse-mouse hybridomas derived from spleen cells. In this study we compared the ability of UF to support growth and antibody production of IgA hybridomas derived from Peyer's patch cells with that of an IgG hybridoma cell line. One IgG (LPC2) and two IgA hybridoma cell lines (RB3 and P2E7) were used as models. The optimal UF concentration for Ig production and cell growth for both the IgA hybridoma RB3 and the IgG hybridomas was 5-10%. Initial plating density was found to be a critical factor for IgA hybridoma cell growth: the IgA hybridomas required a seeding density of at least 70,000 cells/ml to grow compared to 15,000 IgG hybridoma cells/ml (Pakkanen et al., 1992). The addition of small amounts (up to 2%) of FBS in 10% UF supplemented medium did not enhance IgA production or cell growth. RB3 and LPC2 cells seeded at equal density and grown in 10% UF for 8 days attained maximum cell densities at 3-4 days that were 58% (RB3) or 34% (LPC2) lower than those in 10% FBS, but the total amounts of monoclonal antibody produced were 73% and 83%, respectively, of that in 10% FBS. Thus, Ig production per cell was 22-27% higher in 10% UF than in 10% FBS. Hybridoma cells could be cultured for at least 5 weeks without any reduction in growth rates, if medium was partially but not completely replaced twice a week. This suggests that hybridoma cells maintained in UF supplemented medium secrete growth promoting factors. Cells maintained in UF for up to 5 weeks sustained similar monoclonal antibody production rates as in short term culture. These results show that UF can be used as an economical and effective hybridoma culture supplement for the production of both IgG and IgA antibodies.