The Influence of Stimulated Peritoneal Feeder Cells and Mitogens Upon Antibody Secreting Hybridomas

Abstract

Peritoneal exudate cells from mice injected with immunostimulatory agents were evaluated for their ability to promote hybridoma growth. Peritoneal cells from mice receiving peritoneal injections of either Freund's incomplete adjuvant or pristane, seven days prior to harvesting, produced the greatest number of antibody-producing hybridomas. Freund's incomplete adjuvant produced 16 fold more peritoneal cells than unstimulated mice, thus reducing the number of mice needed to supply feeder cells for the hybridoma cultures. In separate experiments a number of B-lymphocyte stimulating lectins and factors were tested for their ability to promote hybridoma growth. 2-mercaptoethanol (25 microM) routinely increased the number of antibody producing hybridomas by 5 to 15 fold. 2-mercaptoethanol had a varying ability to increase the numbers of hybridoma colonies. The cloning efficiency, rate of cell growth and antibody production of hybridoma cell lines, previously produced in the absence of 2-mercaptoethanol could also be increased when this reducing agent was added to the culture medium.