1 Department of Bioengineering and iBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal. Este endereço de email está protegido contra piratas. Necessita ativar o JavaScript para o visualizar..
2 Instituto de Medicina Molecular, Universidade de Lisboa and Hospital de Dona Estefânia, CHLC, Lisbon, Portugal.
3 Erasmus Medical Center iPS facility, Erasmus Medical Center, University Medical Center, Rotterdam, The Netherland.
4 Department of Bioengineering and iBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
5 Institute of Reconstructive Neurobiology, University of Bonn and Hertie Foundation, Bonn, Germany.
6 Instituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal.
7 Department of Neurology, Hospital Sant Joan de Déu (HSJD), Barcelona and CIBER-ER (Biomedical Network Research Centre on Rare Diseases, Instituto de Salud Carlos III), Madrid, Spain.
8 Department of Developmental Biology, Erasmus Medical Center, University Medical Center, Rotterdam, The Netherlands.
- Publicado em Biotechnol J. 2015 Oct;10(10):1578-88
Standardization of culture methods for human pluripotent stem cell (PSC) neural differentiation can greatly contribute to the development of novel clinical advancements through the comprehension of neurodevelopmental diseases.
Here, we report an approach that reproduces neural commitmentfrom human induced pluripotent stem cells using dual-SMAD inhibition under defined conditions in a vitronectin-based monolayer system. By employing this method it was possible to obtain neurons derived from both control and Rett syndrome patients' pluripotent cells. During differentiation mutated cells displayed alterations in the number of neuronal projections, and production of Tuj1 and MAP2-positive neurons. Although investigation of a broader number of patients would be required, these observations are in accordance with previous studies showing impaired differentiation of these cells. Consequently, our experimental methodology was proved useful not only for the generation of neural cells, but also made possible to compare neural differentiation behavior of different cell lines under defined culture conditions. This study thus expects to contribute with an optimized approach to study the neural commitment of human PSCs, and to produce patient-specific neural cells that can be used to gain a better understanding of disease mechanisms.