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. 2021 Jun 16;14(5):2043–2054. doi: 10.1111/cts.13068

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© 2021 The Authors. Clinical and Translational Science published by Wiley Periodicals LLC on behalf of the American Society for Clinical Pharmacology and Therapeutics.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Methotrexate (MTX)‐induced aberrant embryo development after natural fertilization could be partially mitigated by folic acid (FA). (a) Representative images of healthy embryos, which represent the criteria for staging embryos obtained from natural conception. Aberrant embryos observed in the MTX group represented by flattened morulas, blastocysts with atypical inner cell mass, and embryos without zona pellucida. (b) Statistical analysis of the number of embryos after 108 h of mating showing a significant reduction in the total number of embryos of the MTX group, which could not be restored by FA treatment. (c) The percentage of ~ 8 to 16‐cell embryos increased, whereas the blastocyst proportion decreased in the MTX group. FA treatment mainly recovered the proportion of blastocysts. Aberrant embryos were only found in the MTX group. (*p < 0.05, **p < 0.01, Student’s t‐test, n = 6; Scale bars: 50 μm)