Inhibition of Cell Division in Mouse B-Cell Hybridomas: An Overlooked Property of 2-Mercaptoethanol and Its Impact onin vitroAntibody Production

Abstract

Thiol 2-mercaptoethanol (2-ME) has been reported to enhance growth in lymphocytes by various investigators. Some have used 50 μM for growing hybridomas in vitro. Concentrations of 50 and 5 μM in 5% FBS supplemented D-MEM were tested to determine their effects on the growth of 5 monoclonal antibody secreting mouse B cell hybridomas and the myeloma Sp2/O-Ag14. Viability after 24 and 48 h exposure was determined by Trypan blue exclusion. Analysis by one-way ANOVA confirmed that 50 μM 2-ME has a significant negative impact (p<0.05) on hybridoma as well as on myeloma growth, whereas no significant difference (p>0.05) between the control and the 5 μM treatment group was observed after 48 h. Also, no significant difference (p>0.05) in the mortality rates between the control and the treatment groups was found. When combined with the observed protracted doubling time in the 50 μM treatment group, these results indicate that the impact of 2-ME is due to inhibition of cell division. The degree of inhibition was observed to vary between the different hybridomas as well as the myeloma. Although the impact of 2-ME on mitosis has been demonstrated in organisms such as the ciliated protozoan Tetrahymena pyriformis, the yeast Saccharomycess cerevisiae, and the egg of the echinoid the sand dollar Dendraster excentricus, this work demonstrates for the first time that 2-ME impedes the growth of mouse B cell hybridomas. We conclude that adding 2-ME to mouse B cell hybridoma growth media may not be beneficial.