prikaz prve stranice dokumenta Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells
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master's thesis
Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:277341
Kufrin, Vida
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Kufrin, V. (2021). Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:277341

Kufrin, Vida. "Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:277341

Kufrin, Vida. "Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:277341

Kufrin, V. (2021). 'Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:277341

Kufrin V. Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:277341

V. Kufrin, "Generation and characterization of a double reporter cell line for following maturation of hESC-derived β-cells", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:277341

Metadata
TitleGeneration and characterization of a double reporter cell line for following maturation of hESC-derived β-cells
Title (croatian)Uspostavljanje i karakterizacija stanične linije sa sustavom dva reportera kao modela za praćenje sazrijevanja beta-stanica izvedenih iz embrionalnih matičnih stanica
AuthorVida Kufrin
MentorAnthony Galavas (mentor)
MentorInga Urlić (sumentor)
Committee memberInga Urlić (predsjednik povjerenstva)
Committee memberMartina Šeruga Musić (član povjerenstva)
Committee memberDomagoj Đikić (član povjerenstva)
Committee memberRomana Gračan (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-09-28, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Diabetes mellitus is a group of metabolic disorders primarily affecting pancreatic β-cells. Due to the limitations of existing therapy options for insulin-deficient diabetes, great efforts are put into developing cell-based replacement therapies utilizing human pluripotent stem cells. However, existing differentiation protocols, based on developmental principles in vivo, often result in the generation of cells that are not fully mature and functional. To improve and optimize existing protocols for hPSC reprogramming, we developed a novel H1 INSeGFP/ MAFA-mCherry double reporter cell line using CRISPR-Cas9 technology. Herein, detailed characterization and assessment of differentiation potential of novel cell line is reported. H1 INS-eGFP/MAFA-mCherry reporter line can reproducibly and efficiently differentiate to form β-cells while preserving pancreatic identity likewise paternal unmodified H1 cell line. Furthermore, H1 INS-eGFP/MAFA-mCherry derived β-cells express functionality and maturation related genes and can respond to glucose challenges in vitro. Also, both GFP and mCherry integrated reporter genes are fully functional in novel cell line. Reported findings led to the validation of a novel reporter cell line that will serve as a valuable tool in unraveling novel strategies to achieve a high-efficiency differentiation that presents one more step required for the translation of this approach to the clinic setting.
Abstract (croatian)
Dijabetes je metabolički poremećaj koji primarno pogađa β-stanice gušterače. Budući da su postojeće terapijske opcije za pacijente sa smanjenom razinom inzulina ograničene, velike nade polažu se u stanične terapije temeljene na humanim pluripotentnim matičnim stanicama. Međutim, postojeći protokoli diferencijacije, temeljeni na razvojnim principima in vivo, često rezultiraju generacijom β-stanica koje nisu u potpunosti zrele i funkcionalne. Kako bi se protokoli diferencijacije unaprijedili i optimizirali, razvili smo novu dvostruku reportersku liniju H1 INS-eGFP/MAFA-mCherry koristeći tehnologiju CRISPR-Cas9. U ovom radu prikazana je detaljna karakterizacija i procjena diferencijacijskog potencijala nove stanične linije. Linija H1 INS-eGFP/MAFA-mCherry ima sposobnost efikasne i reproducibilne diferencijacije u β-stanice pritom zadržavajući identitet usporediv s nemodificiranom roditeljskom staničnom linijom H1. Nadalje, H1 INS-eGFP/MAFA-mCherry izvedene β- stanice eksprimiraju gene važne za funkcionalnost i sazrijevanje te imaju sposobnost odgovora na promjene u koncentraciji glukoze in vitro. Također, ugrađeni reporterski geni GFP i mCherry su potpuno stabilni i funkcionalni u ovoj staničnoj liniji. Provedeno istraživanje potvrdilo je ispravnost nove stanične linije koja će biti korištena kao vrijedan alat u dizajnu novih strategija za efikasnu diferencijaciju te predstavljati jedan korak više ka kliničkim ispitivanjima novih terapija za dijabetes.
Keywords
Keywords (croatian)
Languagecroatian
URN:NBNurn:nbn:hr:217:277341
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-09-28
Terms of useLicence In copyright icon
Created on2021-10-08 10:38:33