Comparison of toxicogenomic responses to phthalate ester exposure in an organotypic testis co-culture model and responses observed in vivo

Abstract

We have developed a three-dimensional testicular co-culture system (3D-TCS) which mimics in vivo testes. In this study, transcriptomic responses to phthalate esters (PE’s) were compared in the 3D-TCS with responses in rat testes in vivo. Microarray data from the 3D-TCS and from in vivo testes were used to compare changes in gene expression patterns after exposure to developmentally toxic (DTPE) or developmentally non-toxic (DNTPE) phthalate esters. DTPE treatments clustered separately from DNTPE treatments based on principle components analysis both in vitro and in vivo. Pathway analysis using GO-Elite software showed that terms relating to steroid metabolism or reproductive development were enriched both in vitro and in vivo after DTPE exposure. Processes such as cell cycle, cell proliferation and apoptosis were enriched for DTPE treatments in vitro, but not in vivo. Based on these analyses we concluded that transcriptomic responses in the 3D-TCS reflect key aspects of in vivo phthalate toxicity.

The presence of macrophages and inflammatory responses in an in vitro testicular co-culture model of male reproductive development enhance relevance to in vivo conditions

“…Treatment with a variety of toxicants has indicated that the 3D-TCS captures multiple modes of action for testicular toxicity. These include activation of cell death pathways (cadmium) as well as impacts on testosterone production and steroidogenic gene expression (phthalates) (Yu et al 2008, 2009, Wegner et al 2014, Harris et al 2015). …”

From the Cover: An Animal-Free In Vitro Three-Dimensional Testicular Cell Coculture Model for Evaluating Male Reproductive Toxicants

“…Self-renewal and progeny production of spermatogonia are controlled by the neighboring differentiated cells and extracellular matrix (ECM), known as substrate in vivo niches, while Sertoli cells are required for successful differentiation of germ cells in vitro culture systems (Griswold, 1998). The ECM Matrigel-based primary testicular cell model was reported to form a testicular-like multilayered architecture that mimics in vivo characteristics of seminiferous tubules (Harris et al, 2015;Wegner et al, 2013Wegner et al, , 2014Yu et al, 2005Yu et al, , 2008Yu et al, , 2009. As previously demonstrated, this model differentially responds to various phthalate exposures with changes in several cellular pathways, showing sensitive response to testicular toxicity (Harris et al, 2015;Yu et al, 2009).…”

High-Content Analysis Provides Mechanistic Insights into the Testicular Toxicity of Bisphenol A and Selected Analogues in Mouse Spermatogonial Cells

“…There is thus an urgent need to develop effective and practical tools for early screening of chemicals with potential adverse effects using highthroughput, low-cost methodologies that ensure high predictability of human biological responses (Parks Saldutti et al, 2013). Recently, in vitro testicular models have been introduced to environmental chemical safety assessment to improve the predictability of chemical-induced testicular toxicity in human and elucidate mechanisms of chemical toxicity (Harris et al, 2015;Hofmann et al, 2005a;Parks Saldutti, et al, 2013;Wegner et al, 2013Wegner et al, , 2014Yu et al, 2003Yu et al, , 2005Yu et al, , 2009). Some of these in vitro models form a testicular-like multilayered architecture that mimics in vivo characteristics of seminiferous tubules (Yu et al, 2003(Yu et al, , 2005.…”