Metode reprogramiranja somatskih celic

Osovnikar, Pina (author), Ogorevc, Jernej (mentor)

Nekoč je veljalo, da je diferenciacija celic dokončna in da se diferencirane celice ne morejo več spremeniti v noben drug celični tip. Gurdon je s poskusi na žabah že leta 1962 pokazal, da je mogoče iz jedra somatske celice, s prenosom le-tega v izpraznjeno jajčno celico, pridobiti celoten nov organizem. To je bil prvi dokaz, da diferenciacija celic ni dokončna in da je diferencirane celice mogoče reprogramirati do pluripotentnega stanja. S kloniranjem ovce Dolly smo dobili dokaz, da je reprogramiranje celic mogoče tudi pri sesalcih. Bistveno prelomnico na področju je predstavljalo odkritje Yamanake in Takahashija, ki sta leta 2006 odkrila, da je diferencirane somatske celice možno reprogramirati v pluripotentno stanje z vnosom le štirih transkripcijskih dejavnikov (inducirane pluripotentne matične celice – iPSC). Od takrat so se razvile številne metode za reprogramiranje somatskih celic, ki za vnos DNA zaporedja za transkripcijske dejavnike povezane s pluripotenco, uporabljajo retrovirusne in lentivirusne vektorje, linearno DNA, transpozonske vektorje, adenovirusne vektorje, plazmidne vektorje, ali pa temeljijo na direktnem vnosu rekombinantnih proteinov, mRNA, miRNA, komponent sistema CRISPRa ali majhnih molekul. V diplomskem delu je narejen celosten pregled obstoječih metod reprogramiranja, z opisom njihovih prednosti in slabosti. Pregledna tabela, ki predstavlja sintezo rezultatov, nam lahko služi za izbiro metode reprogramiranja celic, ki je optimalna glede na namen nadaljne rabe reprogramiranih celic.

• Language: Slovenian
• Keywords: matične celice, metode reprogramiranje, inducirane pluripotentne matične celice
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [P. Osovnikar]
• Year: 2019
• PID: 20.500.12556/RUL-110388
• UDC: 602.64:602.9(043.2)
• COBISS.SI-ID: 9369465
• Publication date in RUL: 14.09.2019

OSOVNIKAR, Pina, 2019, Metode reprogramiranja somatskih celic [online]. Bachelor’s thesis. P. Osovnikar. [Accessed 3 June 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=110388

Secondary language

• Language: English
• Title: Methodes for somatic cell reprogramming

Abstract

It was believed that cell differentiation process is irreversible and that terminally differentiated cells could not be transformed into any other cell type. With experiments on frogs in 1962, Gurdon showed that it is possible to obtain a whole new organism from the nucleus of somatic cell by transferring it to the enucleated oocyte. This was the first proof that cell differentiation is reversible and that differentiated cells can be reprogrammed to the pluripotent state. By cloning Dolly the sheep evidence that cell reprogramming is also possible in mammals was obtained. An important further breakthrough in the field of cell reprogramming was the discovery of Yamanaka and Takahashi, who in 2006 discovered that differentiated somatic cells could be reprogrammed into pluripotent state by forced expression of only four transcription factors (induced pluripotent stem cells - iPSC). Since then, several reprogramming methods have been developed, which use different strategies for the insertion of pluripotency associated transcription factors, either by DNA sequences insertion (retroviral and lentiviral vectors, linear DNA, transposonic vectors, adenoviral vectors, plasmid vectors) or direct input of recombinant proteins, mRNA, miRNA, components of the CRISPR system or small molecules. In the thesis, a comprehensive review of existing reprogramming methods was made and their advantages and disadvantages presented. The overview of the reprogramming methods is presented in a table that may be helpful in selecting the optimal method considering further applicative use of reprogrammed cells.

• Keywords: stem cells, reprogramming methodes, induced pluripotent stem cells