Assessment of the impact of direct in vitro PFAS treatment on mouse spermatozoa

Reprod Fertil. 2024 Mar 8;5(1):e230087. doi: 10.1530/RAF-23-0087. Print 2024 Jan 1.


Abstract: Poly- and per-fluoroalkyl substances (PFAS) are synthetic environmentally persistent chemicals. Despite the phaseout of specific PFAS, their inherent stability has resulted in ubiquitous and enduring environmental contamination. PFAS bioaccumulation has been reported globally with omnipresence in most populations wherein they have been associated with a range of negative health effects, including strong associations with increased instances of testicular cancer and reductions in overall semen quality. To elucidate the biological basis of such effects, we employed an acute in vitro exposure model in which the spermatozoa of adult male mice were exposed to a cocktail of PFAS chemicals at environmentally relevant concentrations. We hypothesized that direct PFAS treatment of spermatozoa would induce reactive oxygen species generation and compromise the functional profile and DNA integrity of exposed cells. Despite this, post-exposure functional testing revealed that short-term PFAS exposure (3 h) did not elicit a cytotoxic effect, nor did it overtly influence the functional profile, capacitation rate, or the in vitro fertilization ability of spermatozoa. PFAS treatment of spermatozoa did, however, result in a significant delay in the developmental progression of the day 4 pre-implantation embryos produced in vitro. This developmental delay could not be attributed to a loss of sperm DNA integrity, DNA damage, or elevated levels of intracellular reactive oxygen species. When considered together, the results presented here raise the intriguing prospect that spermatozoa exposed to a short-term PFAS exposure period potentially harbor an alternate stress signal that is delivered to the embryo upon fertilization.

Lay summary: PFAS are synthetic chemicals widely used in non-stick cookware, food packaging, and firefighting foam. Such extensive use has led to concerning levels of environmental contamination and reports of associations with a spectrum of negative health outcomes, including testicular cancer and reduced semen quality. To investigate the effects of PFAS on male reproduction, we incubated mouse sperm in a cocktail of nine PFAS at environmentally relevant concentrations before checking for a range of functional outcomes. This treatment strategy was not toxic to the sperm; it did not kill them or reduce their motility, nor did it affect their fertilization capacity. However, we did observe developmental delays among pre-implantation embryos created using PFAS-treated sperm. Such findings raise the intriguing prospect that PFAS-exposed sperm harbor a form of stress signal that they deliver to the embryo upon fertilization.

Keywords: embryo development; environmental contaminants; male fertility; male infertility; male reproduction; perfluoroalkyl and polyfluoroalkyl substances (PFAS); spermatozoa; toxicants.

MeSH terms

  • Animals
  • DNA / pharmacology
  • Fluorocarbons* / toxicity
  • Male
  • Mice
  • Neoplasms, Germ Cell and Embryonal*
  • Reactive Oxygen Species / pharmacology
  • Rodent Diseases*
  • Semen
  • Semen Analysis / veterinary
  • Spermatozoa / physiology
  • Testicular Neoplasms* / veterinary


  • Reactive Oxygen Species
  • DNA
  • Fluorocarbons

Supplementary concepts

  • Testicular Germ Cell Tumor