Use of Primary Mouse Embryonic Fibroblasts in Developmental Toxicity Assessments.

Abstract

Mouse embryonic fibroblasts (MEFs) are commonly collected as a means to maintain the culture and growth of embryonic stem cells (ESCs). However, their utility can extend well beyond their use exclusively in ESC culture. With collection from various transgenic mouse models, use of MEFs may serve as a more simplistic means to reconstitute in vivo/in utero toxicological assessments in an in vitro format for evaluation of function of specific proteins during toxic insults. The ease of collection, rapid growth kinetics, and large-scale expansion to perform multiple, high-throughput experiments are just some of the advantages of MEF use. Here, we describe procedures for successful MEF isolation and culture. As an example of MEF utility, we use MEFs collected form wild-type (WT) and Nrf2 knockout mice. After collection, MEFs were pretreated with the Nrf2 activator, dithiol-3-thione (D3T; 10μM) for 12h, and then treated with either hydrogen peroxide (0-2000μM) or mercury (0-100μM) for another 24h. Viability was measured via MTT assay after 24h of treatment.