Reprod. Nutr. Dev. 42 (2002) 317-326
DOI: 10.1051/rnd:2002028

Neural differentiation of mouse embryonic stem cells grown in monolayer

Jirí Pacherník^{a, b}, Milan Esner^b, Vítezslav Bryja^{a, c}, Petr Dvorák^{a, c, b} and Ales Hampl<sup>a, c, b

a</sup>  Center for Cell Therapy and Tissue Repair, Charles University, Vúvalu 84, 150 06 Prague 5, Czech Republic
^b  Laboratory of Molecular Embryology, Mendel University Brno, Zemedelská 1, 613 00 Brno, Czech Republic
^c  Department of Molecular Embryology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Vídenská 1083, 142 20 Prague, Czech Republic

(Received 26 February 2002; accepted 24 June 2002)

Abstract
To drive neural differentiation of mouse embryonic stem (ES) cells, various culture protocols have been previously developed that all require the formation of embryoid bodies, usually combined with a treatment by all-trans retinoic acid (aRA). Here, we investigated whether or not neural differentiation can also occur in a simplified monolayer culture. Mouse ES cells were plated in serum-containing DMEM media with and without aRA and grown under these conditions for 2 days. Then, the cells were transferred to fresh serum-containing DMEM media and/or to serum-free DMEM/F12 media supplemented with a mixture of insulin, transferrin, selenium, and fibronectin (ITSF) for further culture. The changes in cell morphology and in the expression of selected molecular markers were monitored. Generally, in contrast to all the others, the protocol consisting of a 2-day culture in serum-containing DMEM followed by continuous exposure to the ITSF supplement in DMEM/F12 drove a vast majority of ES cells to generate phenotypic signs of neural lineage. Altogether, neural differentiation can be achieved in vitro without the step involving the formation of embryoid bodies as well as the treatment by aRA.

Key words: embryonic stem cells / neural differentiation

Correspondence and reprints: Ales Hampl
    e-mail: hampl@mendelu.cz

© INRA, EDP Sciences 2002