magistrsko delo
Ana Pleterski (Author), Gregor Majdič (Mentor), Gregor Majdič (Thesis defence commission member), Mojca Narat (Thesis defence commission member), Simon Horvat (Thesis defence commission member)

Abstract

Matične celice pridobljene iz kadavrov predstavljajo obetajoč vir teh celic za različne raziskave kot tudi za regenerativno medicino, sploh kadar je njihovo pridobivanje iz živega darovalca omejeno oz. težavno. V naši raziskavi nas je zanimalo ali po 3, 6 in 9 dneh po smrti še lahko pridobimo celice iz posmtnega tkiva miši. Uporabili smo štiri različna tkiva in sicer, mišično in maščobno tkivo ter tkivo hipokampusa in vohalnih betičev. Prav tako smo preverjali vpliv temperature in način shranitve tkiva na pridobitev celic ter s pomočjo RT-qPCR preverjali izražanje določenih genov, ki so značilni za posamezno razvojno stopnjo celic. Ugotovili smo, da ima temperatura bistven vpliv, saj smo uspeli pridobiti celice le pri 4°C, medtem ko pri sobni temperaturi ne. Vohalni betiči so se izkazali kot tkivo, kjer smo v največ primerih uspeli pridobiti celice, najslabša rast pa je bila pri maščobnem tkivu. Pri nobenem tkivu nismo uspeli pridobiti celic 9 dni po smrti. Prav tako smo ugotovili, da način shranitve tkiva nima bistvenega vpliva. Analiza izražanja genov je pokazala zelo nizko izraženost preiskovanih genov. Izražali so se tako geni za celice, ki še niso končno diferencirane, kot za tiste, ki so. Eni izmed najpogosteje izraženih genov so bili Nanog, Cd34, Sox 7, Eno1, Msln, Gad2, Ptcra, Gad1, Gfap, Alb, Sftpb, G6pc in Tat. Ker ni bilo tipičnega vzorca izraženosti, ne moremo potrditi pluripotentnosti pridobljenih celic.

Keywords

biotehnologija;matične celice;izražanje genov;karakterizacija celic;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL VF - Veterinary Faculty
Publisher: [A. Pleterski]
UDC: 602.9:591.81:576.36/.38(043.2)
COBISS: 8811129 Link will open in a new window
Views: 761
Downloads: 242
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Other data

Secondary language: English
Secondary title: Isolation and characterization of cells from dead mouse tissue
Secondary abstract: Cadavers represent a uniqe and promising source of stem cell which are usefull for many different researchs as well as regenerative medicine. This is importaint especially when there is limited number of donors, small amount of material and difficulties associated with purification. In our research we investigated if we can obtain cells from post mortem mouse tissue 3, 6 and 9 days after death. We used tissue from hippocampus, olfactory bulb, muscle and adipose tissue. We also investigated the inpact of temperature of storage and the form of storage on cell acquisition. In these cells we further analyzed gene expression of 84 key genes for cellular differentiation with RT-qPCR. We found out, that storage temperature has significant impact on acquisition of cells, because we were able to obtain cells only where tissue was stored at 4°C and not on room temperature. It turned out, that tissue from olfactory bulb gave the most positive results with cell growth and the poorly growth was in adipose tissue. We weren't able to obtain cells from any tissue 9 days after death and our results show that form of tissue storage hasn't got any significant impact. Analysis of gene expression has shown very low expression. There were expressed both, genes for terminal differentiated cells as well for progenitor cells. Most commonly expressed genes were Nanog, Cd34, Sox 7, Eno1, Msln, Gad2, Ptcra, Gad1, Gfap, Alb, Sftpb, G6pc and Tat. Since there was no typical pattern of expression we can not confirm the pluripotency of the cells we had obtain.
Secondary keywords: biotechnology;stem cells;gene expression;cell characterization;
Type (COBISS): Master's thesis/paper
Study programme: 0
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Pages: IX, 45 f., [8] f. pril.
ID: 10910807