Vizualizacija mitohondrija u matičnim stanicama

master's thesis

• Title (english): Visualization of mitochondria in stem cells
• Author: Sebastijan Spajić
• Mentor: Dinko Mitrečić (mentor)
• Committee member: Marija Ćurlin (predsjednik povjerenstva)
• Committee member: Ana Katušić (član povjerenstva)
• Committee member: Dinko Mitrečić (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; (Department of Histology and Embryology); Zagreb
• Defense date and country: 2019-07-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Basic Medical Sciences; Cytology, Histology and Embryology
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Basic Medical Sciences; Neuroscience

Abstract

Mitohondriji su stanični organeli zaduženi za proizvodnju ATP-a, reguliranje unutarstanične koncentracije Ca2+ i slobodnih kisikovih radikala te imaju važnu ulogu u intrinzičnom putu aktivacije apoptotičkih mehanizama. Razvojem biologije matičnih stanica dugo se zapostavljao utjecaj mitohondrija na matične stanice te se tek u novije vrijeme istražuje utjecaj mitohondrija na potentnost, proliferaciju i stupanj diferenciranosti matičnih stanica. Kako bi pratili utjecaj mitohondrija na matične stanice važno je uspostaviti metodu njihove vizualizacije. Cilj ovoga istraživanja je bio vizualizirati mitohondrije u oralnim matičnim stanicama čovjeka i živčanim matičnim stanicama miša uz pomoć komercijalne boje MitoTracker. Praćen je utjecaj normoksijskih i hipoksijskih uvjeta na oblik i količinu mitohondrija u živčanim matičnim stanicama miša. Jedna skupina stanica je bila cijelo vrijeme u normoksiji do trenutka fiksacije. Druga skupina stanica je bila 24 sata u hipoksijskim uvjetima, a zatim je provela 6 te 24 sata u normoksijskim uvjetima do trenutka fiksacije. U objema populacijama matičnih stanica uspjeli smo uz MitoTracker® Red CMXRos vizualizirati mitohondrije i odrediti oblik mitohondrija. Ustvrdili smo da uzgoj živčanih matičnih stanica u normoksijskim uvjetima ne mijenja mitohondrijsku dinamiku, dok uzgoj živčanih matičnih stanica prvo u hipoksijskim, a zatim u normoksijskim uvjetima mijenja mitohondrijsku dinamiku.

Abstract (english)

Mitochondria are cell organelles in charge of ATP production, regulation of intracellular Ca2+ concentration and reactive oxygen species and they have an important role in the intrinsic pathway for activation of apoptotic mechanisms. Mitochondria play a key role in all cells, including stem cells. The development of stem cell biology has long neglected the effects of mitochondria on stem cells, and only recent studies have investigated the influence of mitochondria on potency, proliferation and the degree of differentiation of stem cells. In order to monitor the mitochondrial influence on stem cells, it is important to visualize the mitochondria themselves. The aim of this study was to visualize mitochondria in human oral mucosa stem cells and mouse neural stem cells using the commercial dye MitoTracker. The influence of normoxic and hypoxic conditions on the shape and amount of mitochondria in mouse neural stem cells was observed. One group of stem cells was in normoxia until they underwent fixation. Another group of cells spent 24 hours in hypoxia. After that, one of the subgroups spent 6 hours and another one 24 hours in normoxia until they were fixated. Using MitoTracker® Red CMXRos, we succeeded in visualizing and defining the shape of mitochondria in both stem cell populations. We have found that, under normoxic conditions, the mitochondrial dynamics of neural stem cell does not change, whereas if the neural stem cells are first put under hypoxic and then under normoxic conditions, their mitochondrial dynamics does change.

• Keywords: mitohondriji; matične stanice; mitotracker; hipoksija; normoksija
• Keywords (english): mitochondria; stem cells; mitotracker; hypoxia; normoxia
• Language: croatian
• URN:NBN: urn:nbn:hr:105:846665
• Study programme: Title: Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-04-28 09:46:42