ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

and HistoPARK (CZ.1.07/2.3.00/20.0185). LK was supported by the European Regional
Development Fund - Project FNUSA-ICRC (No. CZ.1.05/1.1.00/02.0123).
References
Chandel, N.S., McClintock, D.S., Feliciano, C.E., Wood, T.M., Melendez, A., Rodriguez,
A.M., Schumacker, P.T.: Reactive oxygen species generated at mitochondrial complex III
stabilize hypoxia-inducible factor-1α during hypoxia: a mechanism of O2 sensing. – J Biol
Chem 275: 25130-25138, 2000.
Finkel, T.: Signal transduction by reactive oxygen species. – J Cell Biol 194 (1): 7-15, 2011.
Wang G. L., Jiang B. H., Rue E. A., Semenza G. L.: Hypoxia-inducible factor 1 is a basichelix-loop-helix-PAS
heterodimer regulated by cellular O2 tension. – Proc Natl Acad Sci
USA 92: 5510–5514, 1995.
Wouters, B. G. and Koritzinsky, M.: Hypoxia signalling through mTOR and the unfolded
protein response in cancer.: – Nat Rev Cancer. 8: 851-64, 2008.
P87. ROLE OF HIF-1Α IN SELFRENEW AND DIFFERENTIATION OF NEUROSPHERES
DERIVED FROM EMBRYONIC STEM CELLS
Pánská V. 1 , Večeřa J. 1 , Kubala L. 2,3 , Pacherník J. 1,3
1
Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech
Republic; veronika.panska@centrum.cz
2
Institute of Biophysics ASCR v.v.i., Brno, Czech Republic
3
ICRC - CBCE, FNUSA, Brno, Czech Republic
Hypoxia inducible factor 1α (HIF-1α) is a main factor which responds to hypoxia
and regulates adaptation to hypoxic conditions. HIF-1α 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-1α deficient embryonic stem cells (ES) and
NSCs dissected from embryonic brain (Hitoshi et al., 2004). We monitor influence of
HIF-1α 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-1α
supports selfrenewal of NSCs and knockout of HIF-1α enables neurospheres to express
more neural markers. Moreover, gene expression of Hes1 and Hes5 transcription factors
is affected by HIF-1α knockout which poits to interaction of HIF-1α and Notch pathways
in selfrenewal abilities of NSC.
Acknowledgements
192 Analytical Cytometry VII