In vitro testicular toxicity of environmentally relevant endocrine-disrupting chemicals: 2D vs. 3D models of prepubertal Leydig TM3 cells

• Published in: Environmental toxicology and pharmacology Vol. 93; p. 103869
• Main Authors: Sychrová, Eliška, Yawer, Affiefa, Labohá, Petra, Basu, Amrita, Dydowiczová, Aneta, Virmani, Ishita, Babica, Pavel, Sovadinová, Iva
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2022; Elsevier Science Ltd
• Subjects: 3D in vitro models; Animals; Benzo(a)pyrene; Benzo(a)pyrene - toxicity; Cell culture; Chemicals; Cytotoxicity; Disruption; Endocrine disruptors; Endocrine Disruptors - chemistry; Endocrine-disrupting chemicals; Fluoranthene; Hazard assessment; Health hazards; Leydig Cells; Lipotoxicity; Male; Males; Methoxychlor; Mice; Pyrene; Reproduction; Reproductive health; Reproductive system; Reproductive toxicity; Spheroids; Steroidogenesis; Testes; Testis; Three dimensional models; Toxicity; Triclocarban; Triclosan; 3D in vitro models; Lipotoxicity; Endocrine-disrupting chemicals; Reproductive toxicity; Spheroids; Leydig cells
• ISSN: 1382-6689; 1872-7077
• DOI: 10.1016/j.etap.2022.103869

The testis is a priority organ for developing alternative models to assess male reproductive health hazards of chemicals. This study characterized a 3D in vitro model of murine prepubertal Leydig TM3 cells with improved expression of steroidogenesis markers suitable for image-based screening of testicular toxicity. This 3D scaffold-free spheroid model was applied to explore the impact of prototypical endocrine-disrupting chemicals (EDCs) and environmental reprotoxicants (benzo[a]pyrene, 2- and 9-methylanthracenes, fluoranthene, triclosan, triclocarban, methoxychlor) on male reproductive health. The results were compared to the male reprotoxicity potential of EDCs assessed in a traditional monolayer (2D) culture. The testicular toxicity was dependent not only on the type of culture (2D vs. 3D models) but also on the duration of exposure. Benzo[a]pyrene and triclocarban were the most active compounds, eliciting cytotoxic effects in prepubertal Leydig cells at low micromolar concentrations, which might be a mechanism contributing to their male reprotoxicity. [Display omitted] •testicular toxicity of EDCs was dependent on the type of culture: 2D vs. 3D.•EDCs exhibited distinct time-response toxicity profiles.•B[a]P and TCC were the most active EDCs and toxic at low micromolar concentrations.•B[a]P and TCC induced lipid accumulation in Leydig TM3 cells.