Adhezija modelnih celic na kovinske elektrode za uporabo v biosenzoriki

magistrsko delo

Ajda Sušnik (Author), Urban Bren (Mentor), Martin Rozman (Co-mentor)

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;magistrske naloge;

Data

Language:	Slovenian
Year of publishing:	2020
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[A. Sušnik]
UDC:	602.1:53.082.9(043.2)
COBISS:	47526915
Views:	486
Downloads:	77
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Adhesion of model cells to metal electrodes for use in biosensoric applications
Secondary 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.
Secondary keywords:	cell adhesion;HepG2 cells;metal electrodes;impedance biosensor;
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	XI, 64 f.
ID:	12258010