This year's training was the extension of the work I had done  previously on establishing Danio rerio testis tissue culture. Establishment and optimization were not completed during my first stay and there was a plan to finalize it. The procedure is of interest both for my lab as well as for the lab of the Department of Bioanalytical Ecotoxicology because such system could be used for testing endocrine disrupting substances on the level of testicular steroidogenesis in fish. As such system was also something new for UFZ lab, dr Scholz made suggestion that we could benefit if I make a short visit to the Department of Biology, division of Endocrinology and Metabolism, Utrecht University, Netherlands to visit dr. Rüdiger W. Schulz’s lab from where actually this procedure originates in a form we wanted it to perform. Visit fulfilled our expectation, I gained some specific skills necessary to successfully conduct  testis tissue culture experiments. In Leipzig I was able to apply knowledge, culture conditions were now set correctly and I could say establishment of the culture succeeded. As it was obvious it was working, I started with my experiments, testing toxicity of hexabromocyclodoedecane (HBCDD) – a brominated flame retardant and analyzed its toxicity upon gene expression of enzymes involved in fish steroidogenesis using Real-Time PCR.
The technique of culturing is somewhat unique in a way it is performed. For the set up of experiment, one needs to make cylinders of agarose gels in a 48-well plate, prepare nitrocellulose membrane cuts and to place it together to 24-well plate for presoaking over night in medium containing test substances. Next day, isolated testes from fish are put on the top of the membranes which lays on the top of the gels. Testes should not be covered with medium because they need good aeration, medium is below the level of the testes. If conditions are set correctly, testes can survive up to 7 days in such environment, possible even more. For the purpose of inspecting steroidogenic system, 1-2 days are maximum period of culturing in a way to have this process working outside of living organism Method might have its weaknesses, but with right optimization it can be very interesting because it is relatively new method and we plan to introduce the technique to our lab in Novi Sad.
I’m grateful  for the opportunity to work and learn at dr Scholz’s lab in Leipzig as well as to visit and learn at dr Schulz’s lab in Utrecht. I find it very useful for me personally as well as for our lab LECOTOX. It was also  wonderful experience in a way to meet  old and new friends and see how it looks like to spend summer in Germany!




Svetlana Fa

University of Novi Sad, Faculty of Sciences
Department for Biology and Ecology
LECOTOX
D. Obradovica Sq. 2
21000 Novi Sad
tel. +381 21 485 2675
fax. +381 21 450 620
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