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doctoral thesis
IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI
Zagreb: University of Zagreb, Faculty of Science, 2022. urn:nbn:hr:217:213679
Bokulić, Ana
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Bokulić, A. (2022). IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI (Doctoral thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:213679

Bokulić, Ana. "IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI." Doctoral thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:213679

Bokulić, Ana. "IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI." Doctoral thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:213679

Bokulić, A. (2022). 'IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI', Doctoral thesis, University of Zagreb, Faculty of Science, accessed 14 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:213679

Bokulić A. IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI [Doctoral thesis]. Zagreb: University of Zagreb, Faculty of Science; 2022 [cited 2024 May 14] Available at: https://urn.nsk.hr/urn:nbn:hr:217:213679

A. Bokulić, "IZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI", Doctoral thesis, University of Zagreb, Faculty of Science, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:217:213679

Metadata
TitleIZOLACIJA I KARAKTERIZACIJA DIVLJEGA TIPA STANICA MDCK S NISKOM IZRAŽENOŠĆU GENA mdr1 I NJIHOVA PRIMJENA U ISTRAŽIVANJU MEMBRANSKE PROPUSNOSTI
Title (english)ISOLATION AND CHARACTERISATION OF MDCK CELLS WITH LOW EXPRESSION OF mdr1 GENE AND THEIR USE IN MEMBRANE PERMEABILITY SCREENING
AuthorAna Bokulić
MentorAstrid Milić (mentor)
Committee memberDomagoj Đikić (predsjednik povjerenstva)
Committee memberDonatella Verbanac (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2022-03-01, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Universal decimal classification
(UDC)		577 - Biochemistry. Molecular biology. Biophysics
Abstract
U ovoj disertaciji je opisana izolacija klona divljega tipa stanica MDCK s niskom izraženošću gena mdr1 koji kodira P-glikoprotein. Klonovi su izolirani metodom velikog razrjeđivanja, te uspoređeni mjerenjem dvosmjernog transporta prema razini efluksa amprenavira. Klon 79, koji je pokazao najmanji omjer efluksa amprenavira, kao i tri puta manju izraženost mRNA za mdr1 u odnosu na divlji tip (MDCK-WT), odabran je kao klon MDCK s niskim efluksom za daljnje istraživanje. Na standardnom setu spojeva klon 79 pokazao je znatno manji efluks od MDCK-WT, iako je kod nekih spojeva i dalje detektirana ostatna aktivnost P-glikoproteina. Transport 12 makrocikličkih spojeva ispitan je na klonu 79 i nekoliko dodatnih staničnih linija, Caco-2, MDCK-MDR1, MDCK-WT, te MDCK bez ekspresije P-glikoproteina. Nije uočeno značajno poboljšanje permeabilnosti makrolida u staničnim modelima s niskom ili bez izraženosti P-glikoproteina u odnosu na MDCK-MDR1 i MDCK-WT stanični model. Proučen je utjecaj vremenske komponente na permeabilnost makrocikličkih spojeva u staničnom modelu dvosmjernog transporta. Dobiveni rezultati analizirani su multivarijantnim statističkim metodama, te su uspješno razvijeni modeli permeabilnosti i bioraspoloživosti.
Abstract (english)
In this thesis, a clone of wild type MDCK cells with low expression of the mdr1 gene that encodes the efflux pump P-glycoprotein, was isolated using the limiting dilution method. The clones were ranked based on amprenavir efflux level in a bidirectional transport assay. Clone 79 was selected as a low efflux MDCK clone for further investigation since it had the lowest efflux ratio of amprenavir and three fold lower mdr1 mRNA expression in comparison to MDCK wilde type. When profiled using standard compounds, clone 79 had substantially reduced efflux compared to the wild type, although residual P-glycoprotein activity was still observed with some compounds. The transport of 12 macrocyles was tested using clone 79 and several additional cell lines, including Caco-2, MDCK-MDR1, MDCK-WT, and MDCK with no P-glycoprotein expression. No significant improvement in macrolide permeability was observed in cell models with low or no P-glycoprotein expression in comparison to the MDCK-MDR1 and MDCK-WT cells. The influence of the time component on the permeability of macrocyclic compounds in the cellular transport model was further investigated. The obtained results were analyzed by multivariate statistical methods and models of permeability and bioavailability were successfully developed.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:213679
Promotion2022
Study programmeTitle: Biology
Study programme type: university
Study level: postgraduate
Academic / professional title: doktor/doktorica znanosti, područje prirodnih znanosti, polje biologija (doktor/doktorica znanosti, područje prirodnih znanosti, polje biologija)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2022-11-22 11:41:39