An in vitro assay for teratogens with cultures of rat embryo midbrain and limb bud cells.

A short-term in vitro assay for teratogens has been evaluated and shown to have a high predictability (greater than 90%). Cultured cells derived from midbrain (CNS) and limb buds (LB) of 34 to 36 somite rat embryos were exposed to 46 compounds (27 teratogens, 19 nonteratogens) in a blind trial. Rat liver post mitochondrial supernatant fraction plus cofactors were included in the cultures to provide metabolizing enzymes. Differentiation of neurons from CNS cells and chondrocytes from LB cells was measured after 5 days of culture. Inhibition of differentiation (assessed by reduction of number of foci) was the indicator of potential teratogenicity. Variation between experiments was limited. In repeat experiments with two direct-acting teratogens, aldrin and diphenylhyantoin, interexperiment variability was low (coefficient of variation; range 7 to 24%). Of 27 teratogens 25 (93%) and only 2 of 19 nonteratogens (11%) inhibited differentiation (CNS or LB). Inhibition of differentiation in one cell type alone was less predictive (CNS: 85%, LB: 82%).