Zajedničko djelovanje 17α-etinilestradiola i dietilstilbestrola na stanične linije sisavaca i riba

master's thesis

• Title (english): Combined effect of 17α-ethynylestradiol and diethylstilbestrol on mammalian and fish cell lines
• Author: Sanja Sente
• Mentor: Višnja Gaurina Srček (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2012-10-29, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Kulture životinjskih stanica zauzimaju značajno mjesto u biotehnologiji te srodnim znanstvenim područjima kao što su biofarmaceutika, molekularna biologija i toksikologija. Primjenom različitih in vivo i in vitro testova utvrđena je prisutnost velikog broja endokrinih modulatora u okolišu. Endokrini modulatori su spojevi koji oponašaju prirodne hormone i ometaju njihovo djelovanje u organizmu. One tvari koje oponašaju djelovanje estrogenih hormona smatraju se okolišnim estrogenima, a svakodnevno smo im izloženi putem tla, vode, zraka i hrane. 17α-etinilestradiol (EE2) i dietilstilbestrol (DES) su sintetski estrogeni koji imaju negativan utjecaj na reproduktivni sustav ljudi i životinja. Cilj ovog rada je odrediti zajedničko djelovanje EE2 i DES-a na rast i morfološke promjene staničnih linija CHO-K1 (Chinese Hamster Kidney) i CCO (Channel Catfish Ovary). Stanice su tretirane različitim koncentracijma EE2 i DES-a, a citotoksični učinak primjećen je pri svim ispitanim kombinacijama već 24 sata od tretmana. Kod obje stanične linije, najslabije djelovanje EE2 i DES-a zapaženo je pri tretmanu s najnižim koncentracijama (1 μg/mL EE2 i 1 μg/mL DES). Najizraženiji inhibitorni učinak zapažen je kod CHO-K1 stanica tretiranih s 1 μg/mL EE2 i 10 μg/mL DES-a, dok na CCO stanice najjači inhibitorni učinak ima 10 μg/mL EE2 i 1 μg/mL DES-a. Bojanjem CHO-K1 i CCO stanica kristal-ljubičastim i hematoksilin-eozinom, vidljive su promjene u broju stanica i gustoći monosloja tretiranih stanica u odnosu na kontrolne stanice. Zapažene morfološke promjene stanica izloženih zajedničkom djelovanju EE2 i DES-a u skladu su s rezultatima testova citotoksičnosti provedenih primjenom tripan-plavo i Neutral Red metode.

Abstract (english)

Animal cell cultures have significant importance in biotechnology and related scientific fields such as biopharmacy, molecular biology and toxicology. A presence of large number of endocrine disrupting compounds (EDCs) is detected in the environment by using various in vivo and in vitro tests. EDSc are chemical substances which mimics natural hormones and influence their activity in organism. Those compounds which mimic action of estrogenic hormones are called environmental estrogens and humans are being exposed to them daily through soil, water, air and food. 17α-ethynylestradiol (EE2) and diethylstilbestrol (DES) are synthetic estrogens which have negative effect on reproductive system of humans and animals. The purpose of this thesis is to determine a combined effect of EE2 and DES on the growth and morphological changes in CHO-K1 (Chinese Hamster Kidney) and CCO (Channel Catfish Ovary) cell lines. The cells were treated with various concentrations of EE2 and DES and the cytotoxic effect was detected with all of the tested combinations 24 hours after the treatment. In both cell lines a combined effect of EE2 and DES was smallest when cells were treated with lowest concentrations, 1 μg/mL EE2 and 1 μg/mL DES. The highest inhibitory effect was detected in CHO-K1 cells treated with 1 μg/mL EE2 and 10 μg/mL DES, while in CCO cells the highest negative effect was observed after treatment with 10 μg/mL EE2 and 1 μg/mL DES. Staning of treated CHO-K1 and CCO cells with the crystal-violet and haematoxylin-eosin revealed changes in cell number and density of monolayer when compared to control cells. Observed morphological changes in cells exposed to combined effect of EE2 and DES are in agreement with results of cytotoxicity tests obtained by trypan-blue and Neutral Red method.

• Keywords: CCO stanice; CHO-K1 stanice; citotoksičnost; dietilstilbestrol; 17α-etinilestradiol
• Keywords (english): CCO cells; CHO-K1 cells; cytotoxicity; diethylstilbestrol; 17α-ethynylestradiol
• Language: croatian
• URN:NBN: urn:nbn:hr:159:137537
• Study programme: Title: Biotechnology; specializations in: Biochemical Engineering; Course: Biochemical Engineering; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko inženjerstvo (diplomirani inženjer biotehnologije; smjer biokemijsko inženjerstvo)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-17 10:49:40