The embryonic toxicity evaluation of deoxynivalenol (DON) by murine embryonic stem cell test and human embryonic stem cell test models

In the present study, murine embryonic stem cell test (mEST) and human embryonic stem cell test (hEST) models were developed according to protocols of the European Centre for the Validation of Alternative Methods (ECVAM). Different concentrations of DON were administered to mouse embryonic stem cells D3 (mESC-D3), mouse Balb/c-3T3 (3T3) embryo fibroblast cells, and human embryonic stem cells H9 (hESC-H9) for 10 days to detect the 50% inhibitory proliferation concentration (IC50) of mESC-D3 cells, 3T3 cells, and hESC-H9 cells with DON. Differentiation of ESCs was initiated by embryoid body (EBs) formation. EBs were exposed to different concentrations of DONfor 10 days. The expression of cardiomyocyte differentiation gene alpha-myosin heavy chain (α-MHC) was detected by real-time PCR and the 50% inhibition of cardiomyocyte differentiation (ID50) was determined. Based on the values of IC50 and ID50, functions I, II, and III were calculated by three linear discriminant functions in the EST model and the embryotoxicity of DON was described by comparing the three functions. Results of the three endpoints of DON in murine EST were 0.141 μg/ml (IC50 3T3), 0.085 μg/ml (IC50 D3), and 0.110 μg/ml (ID50 D3). The function I (−34.43), function Ⅱ (−18.62), and function III (1.98) were calculated for DON by themEST model. The three endpoints of DON in hEST were 0.13 μg/ml (IC50 3T3), 0.11 μg/ml (IC50 H9), and 0.078 μg/ml (ID50). Function I (−25.97), function Ⅱ (−13.18), and function III (−0.12) were calculated based on IC50 and ID50. Since function III > function II > function I, according to the EST criteria, DON was determined to have strong embryo toxicity both by mEST and hEST. Moreover, the hEST model, which excluded species differences, is suggested to be a more accurate and reliable method for the evaluation of chemical embryotoxicity.