ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.
ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.
ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.
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P83. DIFFERENTIATION OF NEURAL CREST CELLS FROM HUMAN EMBRYONIC STEM<br />
CELLS AND THEIR ODONTOGENIC INDUCTION<br />
Jan Křivánek 1 , Karel Souček 2,3 , Radek Fedr 2 , Eva Švandová 4 , Eva Matalová 4,5 , Aleš<br />
Hampl 1,3<br />
1<br />
Department of Histology and Embryology, Faculty of Medicine, Masaryk University,<br />
Brno, Czech Republic<br />
2<br />
Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech<br />
Republic, v.v.i.<br />
3<br />
Center of Biomolecular and Cellular Engineering, International Clinical Research Center,<br />
St. Anne’s University Hospital, Brno, Czech Republic<br />
4<br />
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech<br />
Republic, v.v.i.<br />
5<br />
Department of Physiology, University of Veterinary and Pharmaceutical Sciences, Brno,<br />
Czech Republic<br />
Neural crest cells (NCC) are transient cell type which differentiates from invaginating<br />
neural plate when two opposite neural folds merge together. These cells are able to<br />
undergo an epithelial-mesenchymal transition, migrate through whole vertebrate<br />
embryo and form many terminal differentiated cell types such as melanocytes,<br />
odontoblasts, Schwann cells, connective tissue cells and many others. From this point<br />
of view NCC represent attractive cell type than can be used in regenerative medicine.<br />
Here we have differentiated human embryonic stem cells (hESCs) into cells possessing<br />
molecular and functional properties of NCC. The key step in the differentiation protocol<br />
was dual inhibition of SMAD signaling in its initial phase followed with FACS-mediated<br />
purification of CD271-positive cells at day 11 of differentiation. The obtained cells were<br />
found, by flow cytometry and immunocytochemistry, to express molecular markers<br />
characteristic of NCC such as HNK1, Sox9, Sox10, and AP2αβ. Importantly, these putative<br />
NCC are also capable of in vitro differentiation into adipogenic cell lineage. When<br />
exposed in vitro to FGF8 these putative NCC up-regulate expression of homeobox genes<br />
that are normally produced early in odontogenesis. Currently, we are investigating the<br />
developmental potential of hESC-derived NCC using chick embryos and also we are<br />
testing the stability of molecular and functional phenotype of these cells when they are<br />
long-term propagated in our modified culture media on dishes coated by poly-L-ornithin<br />
and fibronectin.<br />
This work was supported by: CZ.1.07/2.3.00/20.0185, GAP304/11/1418,<br />
MUNI/A/0962/2012, CZ.1.05/1.1.00/02.0123 and MSM0021622430.<br />
188 Analytical Cytometry VII
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