Uspostava modela eksplantata jetrenih metastaza duktalnog adenokarcinoma gušterače za istraživanja staničnih interakcija

master's thesis

• Title (english): Establishment of a pancreatic ductal adenocarcinoma liver metastases explant model for cell interaction studies
• Author: Iva Šutevski
• Mentor: Marco Gerling (mentor)
• Mentor: Nada Oršolić (sumentor)
• Committee member: Nada Oršolić (predsjednik povjerenstva)
• Committee member: Inga Urlić (član povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Committee member: Petra Peharec Štefanić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2021-06-18, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Duktalni adenokarcinom gušterače (PDAC, engl. pancreatic ductal adenocarcinoma) karakterizira izrazito loša prognoza, te mali napredak u klinici u posljednjih 30 godina. U trenutku postavljanje dijagnoze, većina pacijenata već ima udaljene metastaze, koje su najčešći uzrok smrti. Glavno mjesto metastaziranja gastrointestinalnih tumora je jetra. Unatoč tome, malo se zna o interakcijama tumora i okolnih stanica kod PDAC jetrenih metastaza. Cilj ovog rada je bio uspostaviti model eksplantata PDAC... More jetrenih metastaza za istraživanje staničnih interakcija između tumora i okolnih stanica jetre kroz 1) optimizaciju odabranih uvjeta uzgoja, 2) karakterizaciju vijabilnosti, apoptoze i prisutnosti specifičnih tipova stanica te 3) primjenu modela za snimanje živih stanica. Za to su korišteni precizni tkivni prerezi koji nude nekoliko prednosti: sačuvana struktura i funkcija tkiva, reducirano korištenje životinja u istraživanju i potencijalna primjena u personaliziranoj medicini. MTT test, koji mjeri metaboličku aktivnost, je pokazao da su debljina tkiva od 300 μm te Williamov medij E s 10% seruma, idealni uvjeti za uzgoj, posebice za okolno tkivo jetre. Kroz daljnje eksperimente je pokazano da model eksplantata PDAC jetrenih metastaza zadržava cjelovitost i morfologiju tijekom kultiviranja. U konačnici model je uspješno korišten u svrhu praćenja pojedinačnih stanica na dvo-fotonskom konfokalnom mikroskopu. Less

Abstract (english)

Pancreatic ductal adenocarcinoma (PDAC) has an extremely poor prognosis and little therapeutic progress has been made during the last 30 years. At the time of diagnosis, almost all patients already have distant metastases; the most common cause of death. Even though most gastrointestinal tumours metastasize to the liver, little is known about the tumour-stroma interactions in PDAC liver metastases. This study aims to establish an explant model of PDAC liver metastases for the investigation of... More cellular interactions between tumour and surrounding liver cells. This was performed through 1) optimization of selected culturing conditions, 2) characterisation of viability, apoptosis and persistence of specific cell types, and 3) model application in live-cell imaging. Precise tissue sections, as used in this thesis, offer several advantages: preserved tissue structure and function, reduced use of laboratory animals, and possible application in personalized medicine. The assay quantifying metabolic activity (MTT), revealed 300 μm thick sections cultured in William’s Medium E with 10% serum, as optimal conditions, particularly for the surrounding liver. The optimised model maintained its tissue integrity, morphology and cellular heterogeneity during cultivation. Finally, individual cells inside the explant were successfully tracked using a two-photon confocal microscope. Less

• Keywords: duktalni adenokarcinom gušterače; jetrene metastaze; kultura eksplantata
• Keywords (english): pancreatic ductal adenocarcinoma; liver metastases; explant culture
• Language: croatian
• URN:NBN: urn:nbn:hr:217:446025
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-07-22 10:32:00