• Sperm Chromatin Structure Assay (SCSA)
  Partner/s involved: 30
  Purpose of the test within ReProTect:
  Detect chromatin alterations in sperm
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• Neutral Comet assay in sperm
  Partner/s involved: 30
  Purpose of the test within ReProTect:
  Detect DNA damage in sperm
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• Bovine Spermatozoa Cytoxicity Test
  Partner/s involved: Participant 26, Andrea Galli, ISILS, Italy; Participant 29, Ine D.H. Waalkens-Berendsen, TNO, The Netherlands.
  Purpose of the test within ReProTect:
  Development and application of an in vitro toxicity test on mature spermatozoa using CASA
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• Leydig cell tox test
  Partner/s involved: 2
  Purpose of the test within ReProTect:
  Development of a test to measure toxic effects on Leydig cell function
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• Sertoli cell assay to target compounds with a potential to induce testicular damages
  Partner/s involved: 11 (Pfizer)
  Purpose of the test within ReProTect:
  Provide a test to assess testicular toxicity based on Sertoli toxicity only.
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• Bovine oocytes/embryos for the development of alternative tests
  Partner/s involved: 7
  Purpose of the test within ReProTect:
  The purpose of the test is to verify the suitability of bovine oocytes/embryos for the development of alternative tests for reproductive toxicity.
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• MEA (Mouse Embryo Assay)
  Partner/s involved: 28
  Purpose of the test within ReProTect:
  Building block (pre-implantation embryo development) for the battery for the replacement of the in vivo fertility study (segment I for reprotoxicity assessment).
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• FSH-Signaling and Steroidogenesis in Immortalized Mouse Granulosa Cells
  Partner/s involved: 10
  Purpose of the test within ReProTect:
  We are studying FSH signal transduction (accumulation of intracellular cAMP) as well as progesterone and estradiol synthesis in immortalized granulosa cells in order to detect possible chemical effects on female reproduction.
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• FBA (Follicle BioAssay)
  Partner/s involved: 28
  Purpose of the test within ReProTect:
  Building block (ovarian function) for the battery for the replacement of the fertility study (segment I for reprotoxicity assessment).
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• Human endometrial endothelial cells
  Partner/s involved: 21c
  Purpose of the test within ReProTect:
  To detect chemicals that has effects on the endometrial vasculature and thereby female fertility. The endometrial vasculature is of outmost importance for female bleeding control and fertility. The endometrial endothelial cells are believed to have specific properties compared with other endothelial cells in the body. Cells in primary culture are used since they have retained their functional properties compared with endothelial cell lines where the cells have been modified.
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• Human endometrial explants
  Partner/s involved: 39
  Purpose of the test within ReProTect:
  Detect adverse effects on endometrial receptivity.
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• In vitro test on chorionic villous explants from first trimester human placenta
  Partner/s involved: 41
  Purpose of the test within ReProTect:
  To investigate the effect of chemicals on human trophoblast implantation and development.
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• In vitro test on trophoblast-derived cell line cultures including BeWo, JEG-3, and HTR8/Svneo
  Partner/s involved: 41
  Purpose of the test within ReProTect:
  To set up conditions for analysis of the effect of chemicals on human placenta.
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• Placental transport in human dual perfusion system
  Partner/s involved: 40, 32
  Purpose of the test within ReProTect:
  Study of placental transfer and data for QSAR.
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• Embryonic Stem Cell Test (EST)
  Partner/s involved: 12, 6
  Purpose of the test within ReProTect:
  Assessment of developmental toxicity
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• Rat Postimplantation Whole Embryo Culture (WEC)
  Partner/s involved: 4
  Purpose of the test within ReProTect:
  Detection of embryotoxic chemicals, with specific reference to incorporation of a metabolic activation step
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• Metabolic teratogenicity test (working title)
  Partner/s involved: VHM (Partner 15)
  Purpose of the test within ReProTect:
  Metabolic activation of proteratogens with subsequent in vitro detection of the resulting teratogens
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• Application of array technology to in vitro identification of endocrine disrupters
  Partner/s involved: P8 (ISS), P31 (VITO)
  Purpose of the test within ReProTect:
  To implement high-throughput screens of endocrine disrupters by identifying "signatures" of gene expression changes induced by interferences with nuclear receptor pathways
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• MELN
  Partner/s involved: VITO (P31), Bayer (P13), 3rd laboratory to be defined
  Purpose of the test within ReProTect:
  The purpose of this method is to characterize the estrogen and anti-estrogen activity of test chemicals.
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• Androgen receptor-mediated transactivation assay in PALM cells
  Partner/s involved: VITO
  Purpose of the test within ReProTect:
  The genetically engineered PALM cell line is stably expressing the human androgen receptor (AR) together with the reporter gene luciferase under the control of an androgen responsive element. This means that all steps of androgen-mediated signaling from AR binding to the final synthesis of a (reporter) gene product can take place. Accordingly, PALM cells can be used to assess both androgenic and anti-androgenic potential. In the absence of androgenic stimulus, increased expression of luciferase indicates a test compound's androgenic potential, whereas in the presence of androgenic stimulus a decrease in luciferase expression indicates a test compound's anti-androgenic potential.
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• AR CALUX®
  Partner/s involved: BDS (3), VITO (31), BAYER (13)
  Purpose of the test within ReProTect:
  Because of existing scientific data showing interference with androgen signaling by chemicals in vivo screening assays for androgenicity have been proposed to determine androgenicity of chemicals. Results with the AR CALUX® assay show a good correlation with in vivo androgenic activity of chemicals (Sonneveld et al. 2006, Toxicol. Sci., 89:173-87). The method has discrete advantages that go beyond reduced speed, cost, and animal use, such as the possibility to use human cells and receptors. These may reduce the correlation with the animal model, but will enhance the predictive value of the in vivo model. Therefore, the proposed purpose of the method is to be at least at the level of inclusion as part of a hierarchical approach in the context of regulatory guidelines, and even for full replacement of the in vivo methods.
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• ERalpha CALUX®
  Partner/s involved: BDS (3), VITO (31), BAYER (13)
  Purpose of the test within ReProTect:
  Because of existing scientific data showing interference with estrogen signaling by chemicals in vivo screening assays for estrogenicity have been proposed to determine estrogenicity of chemicals. Results with the ERalpha CALUX® assay show a good correlation with in vivo estrogenic activity of chemicals (Sonneveld et al. 2006, Toxicol. Sci., 89:173-87). The method has discrete advantages that go beyond reduced speed, cost, and animal use, such as the possibility to use human cells and receptors. These may reduce the correlation with the animal model, but will enhance the predictive value of the in vivo model. Therefore, the proposed purpose of the method is to be at least at the level of inclusion as part of a hierarchical approach in the context of regulatory guidelines, and even for full replacement of the in vivo methods.
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• Recombinant human estrogen receptor-α binding assay
  Partner/s involved: No ReProTect partners
  Purpose of the test within ReProTect:
  Cross cutting technology
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• Recombinant rat androgen receptor binding assay
  Partner/s involved: BOKU
  Purpose of the test within ReProTect:
  Cross cutting technology
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