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Title:Adhezija modelnih celic na kovinske elektrode za uporabo v biosenzoriki : magistrsko delo
Authors:ID Sušnik, Ajda (Author)
ID Bren, Urban (Mentor) More about this mentor... New window
ID Rozman, Martin (Comentor)
Files:.pdf MAG_Susnik_Ajda_2021.pdf (3,64 MB)
MD5: 1207CDFD1BC2D5CE31D4FECC09AE92B3
PID: 20.500.12556/dkum/eeaa188a-9a69-4b25-8a02-8f6dbe9b2381
 
Language:Slovenian
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:Namen magistrskega dela je bil s pomočjo različnih metod preučiti adhezijo modelnih celic na površino nerjavnega jekla za uporabo v biosenzoriki. Vse poskuse smo izvedli in vitro na celični liniji humanega karcinoma HepG2. Viabilnost celic HepG2 izpostavljenim različnim koncentracijam aflatoksina B1 (AFB1) in izoniazida (INH) smo proučevali s pomočjo testov citotoksičnosti - test z barvilom resazurin, test vpoja barvila nevtralno rdeče (NR) in test z barvilom Coomassie Brilliant Blue (CBB). Preživetje celic, izpostavljenih različnim koncentracijam AFB1 in INH smo proučevali še z optično mikroskopijo in fluorescenčno mikroskopijo, slednjo na osnovi štetja živih in mrtvih celic na površini nerjavnega jekla s pomočjo programa ImageJ. Z vrstično elektronsko mikroskopijo smo proučevali adhezijo in morfologijo celic, pri čemer smo se osredotočili predvsem na spremembe v obliki, strukturi in velikosti posameznih celic na površini nerjavnega jekla. Elektrokemijska impedančna spektroskopija (EIS) pa je bila uporabljena za spremljanje preživelosti in adhezije celic. Celice HepG2 smo gojili na površini nerjavnega jekla, ki je bila uporabljena kot delovna elektroda v elektrokemijskem sistemu. Preprost in priročen elektrokemični biosenzor na osnovi celic je bil razvit za oceno individualne strupenosti AFB1 in INH na celicah humanega karcinoma HepG2. S pomočjo različnih mikroskopskih tehnik smo tako potrdili, da se celice HepG2 primejo na kovinsko površino. Testiranje elektrod s pritrjenimi celicami pa je pokazalo, da so le te primeren kandidat za uporabo v senzoriki. Zaključki pridobljeni tekom te magistrske naloge potrjujejo, da bi bilo mogoče preiskovane celice uporabiti na jekleni površini kot primer enostavnega in cenovno ugodnega biosenzorja.
Keywords:adhezija celic, celice HepG2, kovinske elektrode, impedančni biosenzor
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[A. Sušnik]
Year of publishing:2020
Number of pages:XI, 64 f.
PID:20.500.12556/DKUM-78373 New window
UDC:602.1:53.082.9(043.2)
COBISS.SI-ID:47526915 New window
NUK URN:URN:SI:UM:DK:5X1PTNLZ
Publication date in DKUM:18.01.2021
Views:1452
Downloads:154
Metadata:XML RDF-CHPDL DC-XML DC-RDF
Categories:KTFMB - FKKT
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:15.12.2020

Secondary language

Language:English
Title:Adhesion of model cells to metal electrodes for use in biosensoric applications
Abstract:Various methods of adhesion of the model cells on stainless steel surface (electrodes) for use in biosensorics were studied. All experiments were performed in vitro on the human HepG2 cancer cell line. With cytotoxicity tests, which included Resazurin dye test, Neutral Red dye adsorption test (NR) and Coomassie Brilliant Blue color test, we investigated the survival of HepG2 cells with the addition of different concentrations of aflatoxin B1 (AFB1) and isoniazid (INH). The survival of cells on the stainless steel surface was qualitatively studied on the basis of counting cells on the surface of metal electrode by fluorescence microscopy, with the Excel and ImageJ programs. Scanning electron microscopy was used to determine the morphology of cells, where we draw attention to the changes in the shape, structure and size of individual cells as well as their attachment and spread on the stainless stell surface. Electrochemical impedance spectroscopy (EIS) was also used to monitor survival and adhesion of the cells. HepG2 cells were grown on the stainless steel surface of the working electrode in the electrochemical system. With the evaluation of individual toxicity of AFB1 and INH on human HepG2 cancer cells, simple cell-based electrochemical biosensor was developed. With various microscopic techniques, adhesion of the HepG2 cells to metal surfaces was observed. Tested electrodes with immobilised cells are suitable for use in sensorics. Investigated cells on the stainless steel surface could be applied in an assembly of a model biosensor.
Keywords:cell adhesion, HepG2 cells, metal electrodes, impedance biosensor


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