Production of antibody by human B cells under serum-free conditions

Abstract

A method is described for growing human B cells in 20 microliter hanging drops in Terasaki plates under serum-free conditions. B cell proliferation and differentiation has a critical dependence for added soybean lipid, while T cell proliferation does not. In this medium, pokeweed mitogen stimulation of separated human B cells induces high levels of immunoglobulin in a T dependent manner. Cells from donors vaccinated with tetanus toxoid and shown to be responders by a conventional culture system, produce high levels of IgG anti-tetanus antibody after antigen stimulation in these serum-free microcultures. The serum free culture system has the novel features of high sensitivity to dose of mitogen or antigen, low background responses and high antibody production with low cell numbers.