Citotoksički učinak lindana na CHO (Chinese Hamster Ovary)-K1 stanice praćeno Trypan Blue i Neutral Red metodama

master's thesis

• Title (english): Cytotoxic effect of lindane on CHO (Chinese Hamster Ovary)-K1 cells followed by Neutral Red and Trypan Blue methods
• Author: Saša Martinčić
• Mentor: Jasna Kniewald (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Toxicology); Zagreb
• Defense date and country: 2006-07-11, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Food Technology

Abstract

Iako je suvremena poljoprivredna proizvodnja nezamisliva bez upotrebe pesticida, njihovi ostaci predstavljaju veliku opasnost za zdravlje ljudi i okoliš zbog kumulacije u vodi, tlu, živim organizmima i hrani. U organizam se pesticidi unose resorpcijom putem kože, ralnim i respiratornim putem. Za utvrđivanje toksičnosti raznih ksenobiotika pa tako i pesticida kao predtestovi koriste se in vitro testovi toksičnosti. U ovom je radu kao test-sustav za praćenje citotoksičkog učinka lindana korištena CHO (Chinese Hamster Ovary)-K1 stanična linija u monosloju uzgojena u Dulbecco MEM mediju uz dodatak 5% NCS-a (Newborn Calf Serum) pri temperaturi od 37C i atmosferi od 95% zraka i 5% CO2. Citotoksički učinak lindana praćen je kolorimetrijskim metodama Trypan Blue i Neutral Red kroz 24, 48 i 72 sata i to u rasponu koncentracija od 20-100 µg lindana/mL medija za uzgoj. IC50 vrijednosti određene su iz grafičkog prikaza ovisnosti % inhibicije rasta stanica o logaritmu upotrebljene koncentracije lindana i primjenom metode Trypan Blue. Određena je IC50 vrijednost od 42,6 µg lindana/mL medija za uzgoj, dok primjenom metode Neutral Red ona iznosi 56 µg lindana/mL medija za uzgoj. Upotrebljene kolorimetrijske metode primjenjive su u ispitivanju citotoksičnosti ksenobiotika u CHO-K1 staničnoj liniji.

Abstract (english)

A modern agricultural production is unthinkable without usage of pesticides. However, their residues are dangerous to human health and enviroment because of their ability of cumulation in water, soil, living organisms and food. They can enter an organisam either by skin, respiratory or gastointestinal system. The toxicity of different xenobiotics can be determined by classical (in vivo) and alternative (in vitro) toxicity tests. In this work CHO (Chinese Hamster Ovary)-K1 cell line in monolayer, cultivated in Dulbecco's MEM supplement with 5 % NCS-a (Newborn Calf Serum) at temperature of 37C in the atmosphere of 95% and 5% CO2, was used as the test-system for determination of lindan toxicity. Cytotoxic effect of lindane was followed by two colorimetric methods: Trypan Blue method and Neutral Red method during 24, 48 i 72 hours, at concentrations range of lindane from 20-100 µg/mL of cultivating medium. IC50 values were calculated from graphical correlation between % inhibition of cell growth and logarithm of applied lindane concentration. Using Trypan Blue method IC50 value is 42,6 µg lindane/mL of cultivating medium while using Neutral Red method IC50 value is 56 µg lindane/mL of cultivating medium. The applied methods can be used for cytotoxicity investigation of different xenobiotics in CHO-K1 cell line.

• Keywords: lindan; kultura animalnih stanica; CHO-K1 stanična linija; citotoksičnost; Trypan Blue i Neutral Red
• Keywords (english): lindane; animal cell culture; CHO-K1 cell line; cytotoxicity; Trypan Blue and Neutral Red methods
• Language: croatian
• URN:NBN: urn:nbn:hr:159:027541
• 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: 2017-09-07 11:33:57