A novel human stem cell‐based biomarker assay for in vitro assessment of developmental toxicity

• Published in: Birth defects research 2022-11, Vol.114 (19), p.1210-1228
• Main Authors: Jamalpoor, Amer, Hartvelt, Sabine, Dimopoulou, Myrto, Zwetsloot, Tom, Brandsma, Inger, Racz, Peter I., Osterlund, Torben, Hendriks, Giel
• Format: Article
• Language: English
• Subjects: Assaying; Biocompatibility; Biomarkers; Biomedical materials; Cardiomyocytes; Cell differentiation; Cytology; developmental toxicity; Differentiation (biology); embryotoxicity; Gene expression; Genes; Hepatocytes; human‐induced pluripotent stem cells; Identification methods; in vitro biomarker assay; In vitro methods and tests; In vivo methods and tests; Morphology; Pluripotency; ReproTracker; Stem cell transplantation; Stem cells; Teratogenicity; Thalidomide; Time dependence; Toxicity; Toxicity testing; Valproic acid
• ISSN: 2472-1727; 2472-1727
• DOI: 10.1002/bdr2.2001

Background Testing for developmental toxicity according to the current regulatory guidelines requires large numbers of animals, making these tests very resource intensive, time‐consuming, and ethically debatable. Over the past decades, several alternative in vitro assays have been developed, but these often suffered from low predictability and the inability to provide a mechanistic understanding of developmental toxicity. Methods To identify embryotoxic compounds, we developed a human induced pluripotent stem cells (hiPSCs)‐based biomarker assay. The assay is based on the differentiation of hiPSCs into functional cardiomyocytes and hepatocytes. Proper stem cell differentiation is investigated by morphological profiling and assessment of time‐dependent expression patterns of cell‐specific biomarkers. In this system, a decrease in the expression of the biomarker genes and morphology disruption of the differentiated cells following compound treatment indicated teratogenicity. Results The hiPSCs‐based biomarker assay was validated with 21 well‐established in vivo animal teratogenic and non‐teratogenic compounds during cardiomyocyte and hepatocyte differentiation. The in vivo teratogenic compounds (e.g., thalidomide and valproic acid) markedly disrupted morphology, functionality, and the expression pattern of the biomarker genes in either one or both cell types. Non‐teratogenic chemicals generally had no effect on the morphology of differentiated cells, nor on the expression of the biomarker genes. Compared to the in vivo classification, the assay achieved high accuracy (91%), sensitivity (91%), and specificity (90%). Conclusion The assay, which we named ReproTracker®, is a state‐of‐the‐art in vitro method that can identify the teratogenicity potential of new pharmaceuticals and chemicals and signify the outcome of in vivo test systems.