Utjecaj matičnih stanica na nekroptozu stanica živčanog sustava in vitro uzrokovanu hipoksijom

doctoral thesis

• Title (english): Influence of stem cells on necroptosis in the cells of nervous system in vitro caused by hypoxia
• Author: Valentina Hribljan
• Mentor: Dinko Mitrečić (mentor)
• Committee member: Željka Krsnik (predsjednik povjerenstva)
• Committee member: Nino Sinčić (član povjerenstva)
• Committee member: Fran Borovečki (član povjerenstva)
• Committee member: Marija Renić (član povjerenstva)
• Committee member: Srećko Gajović (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; Zagreb
• Defense date and country: 2023-03-22, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Basic Medical Sciences; Neuroscience
• Universal decimal classification (UDC): 61 - Medical sciences

Abstract

Hipoksijsko-ishemijska ozljeda mozga u perinatalnom periodu je jedan od najčešćih uzroka cjeloživotnog invaliditeta. U ovom radu smo istražili ulogu stanične smrti po tipu nekroptoze u hipoksijsko-ishemijskom oštećenju stanica živčanog sustava, te ulogu živčanih matičnih stanica u oporavku. Prvi cilj je bio uspostaviti pouzdan model nezrelih stanica živčanog sustava, što je postignuto osmodnevnom diferencijacijom živčanih matičnih stanica miša. Usporedba diferencijacije stanica u ambijentalnoj (20 % kisika) i tkivnoj normokosiji (5 % kisika) je pokazala kako tkivna normoksija povećava izražaj nekih gena vezanih uz matičnost, te onih koji određuju identitet neurona i astrocita. Također smo pokazali kako hipoksija (1 % kisika) u trajanju od 24 h nije dovoljna za pokretanje nekroptoze u nezrelim stanicama živčanog sustava, već je za aktivaciju tog oblika stanične smrti potrebna istovremena deprivacija glukoze (OGD). Budući da inhibitor Necrostatin-1s nije utjecao na nekroptozu nezrelih stanica živčanog sustava, zaključili smo kako je nekroptoza u analiziranom modelu neovisna o kinaznoj aktivnosti RIPK1. Iako matične stanice, niti njihov kondicionirani medij nisu direktno smanjili intenzitet nekroptoze nezrelih stanica živčanog sustava, pokazane su mjerljive dobrobiti: dodane matične stanice su djelomično povratile izražaj biljega neurona TUBB3, dok je kondicionirani medij djelomično povratio izražaj SOX2, jednog od proteina važnog za oporavak živčanog sustava.

Abstract (english)

Hypoxic-ischemic encephalopathy is one of the most common causes of lifelong disability. Here, we have investigated the role of necroptosis, a type of cell death, in hypoxic-ischemic insult of cells of the nervous system, and the role of neural stem cells (NSC) in recovery. The first goal was to establish a model of immature cells of the nervous system, which was achieved through the eight-day differentiation of mouse NSC. A comparison of cell differentiation in ambient (20 % oxygen) and tissue (5 % oxygen) normoxia revealed that tissue normoxia increases the expression of some of the genes involved in stemness and neuronal and astrocytic identity. Since activation of necroptosis in immature cells couldn't be achieved by hypoxia (1 % oxygen) during 24 h, we caused necroptosis by oxygen-glucose deprivation. Since inhibitor Necrostatin-1s didn't influence necroptosis, we have concluded that necroptosis in the analyzed model is not dependent on the kinase activity of RIPK1. Although NSC, nor their conditioned medium didn't directly reduce the intensity of necroptosis, we have discovered other benefits: NSC partly recovered the expression of the neuronal marker TUBB3, while the conditioned medium partly recovered the expression of SOX2, a protein important for recovery of the nervous system.

• Keywords: Nekroptoza; Nekrostatin-1; Kisik; Glukoza; Diferencijacija stanica; Neuralne matične stanice
• Keywords (english): Necroptosis; Necrostatin-1; Oxygen; Glucose; Cell Differentiation; Neural Stem Cells
• Language: croatian
• URN:NBN: urn:nbn:hr:105:243257
• Study programme: Title: Neuroscience; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstvo (doktor/doktorica znanosti, područje biomedicine i zdravstvo)
• Type of resource: Text
• Extent: 104 str.
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2024-03-22
• Created on: 2023-04-06 08:31:41