Introduction: This work adapts a novel ovary culture technique, devised by De Felicis group, for assessing the ovotoxicity of two chemotherapeutic drugs, cisplatin and doxorubicin. Previous work from our laboratory used cultures of intact neonatal ovaries to determine the effects of the two drugs, showing that both cisplatin and doxorubicin are moderately ovotoxic.
Methods: Here, ovaries collected from newborn WT mice were cultured as by De Felici. Briefly, ovaries were fragmented and seeded into the wells of a culture plate with serum containing medium. These were left for 3 days to allow the fragments to attach, with cisplatin and doxorubicin added into the culture on day 7 only, at patient serum concentrations of 0.5 and 1 μg/ml, 0.1 and 0.2 μg/ml respectively. Immediately before and after drug administration, tissue fragments were stained with trypan blue to assess cell viability and follicle health.
Results and discussion: From the total of 152 fragments analysed, the percentage of the growing follicles regarded as morphologically unhealthy by trypan blue staining, was significantly lower in the controls (1.98%) as compared to 22.1% (P<0.05) and 25.2% (P<0.01) seen in those treated with 0.5 and 1 μg/l cisplatin respectively, and 41.0% (P<0.0001) and 44.2% (P<0.0001) seen in those treated with 0.1 and 0.2 μg/ml doxorubicin respectively. The culture method used here offers a quick and easy way to evaluate the effects of toxic drugs on ovarian tissue without the need for either in vivo experimentation or extensive histological analysis.
02 - 04 Sep 2014
World Congress of Reproductive Biology