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Use of two validated in vitro tests to assess the embryotoxic potential of mycophenolic acid

  • Reproductive Toxicology
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An Erratum to this article was published on 29 November 2009

Abstract

Mycophenolate mofetil is a widely used immunosuppressive drug that recently has been categorized as a human teratogen. Animal experiments indicate a teratogenic potential of the drug, but so far, it has not been studied in embryotoxicity in vitro assays. The aim of this study was to evaluate the in vitro embryotoxic potential of mycophenolic acid and investigate the ability of such tests to detect its embryotoxic potential. We used two validated assays: the rat whole embryo culture and the murine embryonic stem cell test. Rat embryos cultured from gestational day 9.5 for 48 h with the drug showed dysmorphogenic development already at a concentration of 250 μg mycophenolic acid/l medium. At concentrations of 750 μg/l and more, all rat embryos exhibited malformations. The main effects were defective yolk sac blood circulation, neural tube defects (open cranial neural pore), malformations of the head with missing eye anlagen and heart anomalies. Moreover, the exposed embryos showed a concentration-dependent decrease in protein content, crown-rump length, number of somites and morphological score. The murine embryonic stem cell test was slightly more sensitive. Proliferation and differentiation of the ES-D3-cells were significantly impaired at concentrations of 31 and 125 μg mycophenolic acid/l medium, respectively. In the differentiation assay, at a concentration of 125 μg mycophenolic acid/l medium and more, the number of wells with differentiated cardiomyocytes significantly decreased. Additionally, a cytotoxicity assay with balb/c 3T3 mouse fibroblasts was used to compare the proliferation and vitality of embryonic cells with adult cells. In the balb/c 3T3 cytotoxicity assay, the number of vital mouse fibroblasts significantly decreased at a mycophenolic acid concentration of 62 μg/l and more. In conclusion, by using the two validated in vitro tests, we showed that mycophenolic acid exhibits a pronounced embryotoxic potential at cytotoxic concentrations. This result from validated in vitro tests is of special interest, because it supports the use of the tests to detect human teratogens.

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Acknowledgments

The authors thank Nicole Müller and Annegret Felies for their excellent technical assistance in this study.

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Authors and Affiliations

  1. Institute of Clinical Pharmacology and Toxicology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Garystr. 5, 14195, Berlin, Germany

    Kathrin Eckardt & Ralf Stahlmann

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  1. Kathrin Eckardt
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  2. Ralf Stahlmann
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Correspondence to Ralf Stahlmann.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s00204-009-0492-1

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Eckardt, K., Stahlmann, R. Use of two validated in vitro tests to assess the embryotoxic potential of mycophenolic acid. Arch Toxicol 84, 37–43 (2010). https://doi.org/10.1007/s00204-009-0476-1

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  • DOI: https://doi.org/10.1007/s00204-009-0476-1

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