Notch i Ikaros u diferencijaciji hematopoetskih stanica

master's thesis

• Title (english): Notch and Ikaros in hematopoietic cell differentiation
• Author: Matea Konjević
• Mentor: Mariastefania Antica (mentor)
• Mentor: Maja Matulić (mentor)
• Committee member: Maja Matulić (predsjednik povjerenstva)
• Committee member: Mariastefania Antica (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2012-02-14, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Hematopoeza je proces nastajanja novih krvnih stanica. Transkripcijski faktori iz porodice Ikaros i puta dojavljivanja Notch reguliraju važne kontrolne točke tijekom hematopoeze koje će odlučiti u kojem će se smjeru stanica diferencirati. U radu je potvrđena i izmjerena ekspresija transkripcijskih faktora iz porodice Ikaros: Ikaros, Helios i Aiolos te puta dojavljivanja Notch: Notch1 i Hes1 u NB4 stanicama prije i poslije diferencijacije ATRA-om. U radu je uspješno uhodan, prilagođen i primijenjen protokol za metodu višestruke lančane reakcije polimerazom (multiplex PCR). Metodom je istovremeno umnoženo 6 odsječaka DNA u uzorku od 2500 stanica. Za multiplex PCR su odabrane početnice te provjerene i testirane na specifičnost i izbjegavanje sekundarnih struktura. Semi-kvantitativni RT-PCR je korišten u svrhu otkrivanja moguće promjene u razinama mRNA u stanicama prije i poslije diferencijacije. Smanjenje u ekspresiji gena Aiolos i Hes1 ukazuje na njihovu moguću ulogu u diferencijaciji i razvoju nezrelih mijeloidnih stanica.

Abstract (english)

Hematopoiesis is the process of formation of new blood cells. Transcription factors from the Ikaros family and Notch signaling pathway regulate important checkpoints during hematopoiesis, which will decide in which direction the cells differentiate. In this work was confirmed and measured expression of transcription factors from the Ikaros family: Ikaros, Helios and Aiolos and Notch signaling pathway: Notch1 and Hes1 in NB4 cells before and after differentiation induced with ATRA. Protocol for the method of multiplex polymerase chain reaction (multiplex PCR) was successfully established, adapted and implemented. Six segments of DNA was simultaneously amplified in a sample of 2500 cells. For multiplex PCR, primers were selected and tested for specificity and to avoid secondary structures. Semi-quantitative RT-PCR was used to detect possible changes in the levels of mRNA in cells before and after differentiation. The reduction in expression of genes Aiolos and Hes1 suggests their possible role in differentiation and development of immature myeloid cells.

• Keywords: transkripcijski faktori; višestruka lančana reakcija polimerazom; semi-kvantitativni RT-PCR; Helios; Aiolos; Hes1
• Keywords (english): transcription factors; multiplex polymerase chain reaction; semi-quantitative RT-PCR; Helios; Aiolos; Hes1
• Language: croatian
• URN:NBN: urn:nbn:hr:217:434380
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-02 08:47:56