Informace o publikaci

Role of HIF-1α in selfrenew and differentiation of neurospheres derived from embryonic stem cells



Rok publikování 2013
Druh Další prezentace na konferencích
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Popis Hypoxia inducible factor 1 alpha (HIF-1a) is a main factor which responds to hypoxia and regulates adaptation to hypoxic conditions. HIF-1a is important for stemness maintenance by stabilizing activated Notch-1 (NICD) and for maintenance of neurogenic niche by upregulating vascular endothelial grow factor (VEGF) (Gustafsson et al., 2005; Covello et al., 2004). These informations are important for our experiments with neural stem cells (NSC). We use NSCs derived from murine wt and HIF-1a deficient embryonic stem cells (ES) and NSCs dissected from embryonic brain (Hitoshi et al., 2004). We monitor influence of HIF-1a knock-out on formation, selfrenewal capacity and transition ability of NSCs by neurosphere cultivation (colony forming assay) and by screening of neural markers using qRT-PCR and WB techniques. We also use flow cytometry for confirmation of neural attributes of our NSCs and for quantifying cells with neural characteristics. In this assay we work with neural markers such as Forse-1 and Pax-6. According to our preliminary results from qRT-PCR and colony forming assay, HIF-1a supports selfrenewal of NSCs and knockout of HIF-1a enables neurospheres to express more neural markers. Moreover, gene expression of Hes1 and Hes5 transcription factors is affected by HIF-1a knockout which poits to interaction of HIF-1a and Notch pathways in selfrenewal abilities of NSC.

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