H295R Steroidogenesis Assay (assigned to external lab)

Test organism

  • Human adrenocortical carcinoma cells

Detectable effects (impact)

  • Changes in steroid hormone concentration (impact on steroidogenesis)

Test principle

  • In vitro assay system for determining changes in the formation of steroid hormones. A human cell line capable of expressing all enzymes of the steroidogenic metabolic pathway is used. Moreover, the cell line has the potential of producing all steroid hormones of the fetal and adult adrenal cortex.
  • The hormone concentration (e.g. testosterone, estradiol) is determined with commercially available kits (e.g. Elisa).

Test duration

  • At least 3 d (exposure time: 48 h)

Relevance

  • In vitro assay system for the identification of receptor-dependent and non-receptor-dependent effects of hormonally active substances.
  • Suitable for determining whether steroidogenesis can be affected by chemicals and environmental samples
  • Tool for investigating the correlations between chemicals and the molecular and biochemical processes of steroidosynthetic pathways

Guidelines and literature

  • ISO 19040-1:2018 Water quality — Determination of the estrogenic potential of water and waste water — Part 3: In vitro human cell-based reporter gene assay
  • Gracia T, Hilscherova K, Jones PD, Newsted JL, Zhang X, Hecker M, Higley EB, Sanderson JT, Yu RMK, Wu RSS, Giesy JP (2006). The H295R system for evaluation of endocrine-disrupting effects. Ecotoxicol. Environ. Saf. 65:293-305.
  • Hecker M, Newsted JL, Murphy MB, Higley EB, Jones PD, Wu R, Giesy JP (2006). Human adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: Hormone production. Toxicol. Appl. Pharmacol. 217:114-124.
  • Hecker M, Giesy JP (2008). Novel trends in endocrine disruptor testing: The H295R Steroidogenesis Assay for identification of inducers and inhibitors of hormone production. Anal. Bioanal. Chem. 390:287-291.