Transferability and inter-laboratory variability assessment of the in vitro bovine oocyte maturation (IVM) test within ReProTect.
The new European chemicals policy for the Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) will most probably impose a dramatic increase in the number of animals required for reproductive toxicity testing. For this purpose, the development and validation of alternative methods is urgently needed in order to reduce the use of laboratory animals. The present study describes the inter-laboratory variability and the transferability assessment of an in vitro test able to identify chemical effects during the process of oocyte maturation in a bovine model. The test was developed/optimised within ReProTect, an integrated research project funded by the European Union, joining together 35 partners with complementary expertise in reproductive toxicology. Eight chemicals with well-known toxic properties were tested (benzo[a]pyrene, busulfan, cadmium chloride, cycloheximide, diethylstilbestrol, ketoconazole, methylacetoacetate, mifepristone/RU-486 and DMSO as solvent) on the in vitro maturation (IVM) assay in two well-trained laboratories using the established Standard Operating Procedures. The statistical analysis demonstrated the concordance of results across the laboratories and the reproducibility of the test. We therefore conclude that the IVM test could advance toward the process of validation as alternative in vitro method that, in combination with additional in vitro tests, can become part of an integrated testing strategy in order to predict chemical hazards on mammalian fertility.
Dipartimento di Scienze Animali, Sezione di Anatomia ed Istologia Veterinaria, Università degli Studi di Milano, Milano, Italy. alberto.luciano@unimi.it
Alberto M
AM
Luciano
Federica
F
Franciosi
Valentina
V
Lodde
Davide
D
Corbani
Giovanna
G
Lazzari
Gabriella
G
Crotti
Cesare
C
Galli
Cristian
C
Pellizzer
Susanne
S
Bremer
Marc
M
Weimer
Silvia C
SC
Modina
D000826:Animal Testing Alternatives; D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D015386:Hazardous Substances / Q000633:toxicity*; D066298:In Vitro Techniques; D007753:Laboratories / Q000592:standards; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development; D009866:Oogenesis / Q000187:drug effects*; D015203:Reproducibility of Results; D018675:Toxicity Tests / Q000379:methods* / Q000592:standards
D015386:Hazardous Substances