title: 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.
abstract: Our 3-dimensional testis co-culture system (3D-TCS) represents a promising model of male reproductive toxicity which captures sensitive processes of male reproductive development and contains the main testes cell types (germ, Leydig and Sertoli cells). Macrophages are another cell type important for testicular function and help to modulate immuno-endocrine processes during testes development. Chemicals such as phthalate esters (PE's) affect macrophage function and testosterone production in the testes in vivo. The aim of this study was to determine whether macrophages were present in the 3D-TCS and investigate responses in our model that may be related to immuno-endocrine functions. We observed consistent expression of the resident macrophage marker ED2 as well as increases in inflammatory cytokines produced by macrophages and testes cells (IL-6, TNF-α and KC/GRO) after exposure to toxic PE's. Pathway analysis of gene expression changes after exposure to PE's showed that IL-6 and TNF-α signaling pathways were enriched after treatment with reproductively toxic, but not non-reproductively toxic phthalates. These results indicate that macrophages and inflammatory processes are captured in the 3D-TCS and that these processes are impacted by exposure to reproductive toxicants. These processes represent a major mode of action for in vivo testis toxicity for a variety of compounds and our novel in vitro model is able to capture toxicant perturbation of immune function.
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States.
authors: Sean
authors: S
authors: Harris
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States; Institute for Risk Analysis and Risk Communication, University of Washington, Seattle, WA 98105, United States.
authors: Sara Pacheco
authors: SP
authors: Shubin
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States.
authors: Susanna
authors: S
authors: Wegner
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States; Institute for Risk Analysis and Risk Communication, University of Washington, Seattle, WA 98105, United States.
authors: Kirk
authors: K
authors: Van Ness
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States.
authors: Foad
authors: F
authors: Green
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States; Institute for Risk Analysis and Risk Communication, University of Washington, Seattle, WA 98105, United States.
authors: Sung Woo
authors: SW
authors: Hong
authors: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, United States; Institute for Risk Analysis and Risk Communication, University of Washington, Seattle, WA 98105, United States. Electronic address: faustman@u.washington.edu.
authors: Elaine M
authors: EM
authors: Faustman
mesh_terms: D000818:Animals; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005854:Germ Cells / Q000187:drug effects / Q000378:metabolism; D007985:Leydig Cells / Q000187:drug effects / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D010795:Phthalic Acids / Q000633:toxicity*; D017207:Rats, Sprague-Dawley; D012098:Reproduction; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism; D013737:Testis / Q000166:cytology*; D059467:Transcriptome / Q000187:drug effects
chemical_list: D016207:Cytokines; D010795:Phthalic Acids
keywords: Cytokines; In vitro models; Inflammation; Macrophages; Phthalate esters; Reproductive toxicity; Testis