Neural commitment of human pluripotent stem cells under defined conditions recapitulates neural development and generates patient‐specific neural cells
Abstract
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 commitment from 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.
Number of times cited according to CrossRef: 8
- Carlos AV Rodrigues, Teresa P Silva, Diogo ES Nogueira, Tiago G Fernandes, Yas Hashimura, Robin Wesselschmidt, Maria Margarida Diogo, Brian Lee and Joaquim MS Cabral, Scalable culture of human induced pluripotent cells on microcarriers under xeno‐free conditions using single‐use vertical‐wheel™ bioreactors, Journal of Chemical Technology & Biotechnology, 93, 12, (3597-3606), (2018).
- Cláudia C. Miranda, Tiago G. Fernandes, Sandra N. Pinto, Manuel Prieto, M. Margarida Diogo and Joaquim M.S. Cabral, A scale out approach towards neural induction of human induced pluripotent stem cells for neurodevelopmental toxicity studies, Toxicology Letters, 10.1016/j.toxlet.2018.05.018, 294, (51-60), (2018).
- N Mellios, D A Feldman, S D Sheridan, J P K Ip, S Kwok, S K Amoah, B Rosen, B A Rodriguez, B Crawford, R Swaminathan, S Chou, Y Li, M Ziats, C Ernst, R Jaenisch, S J Haggarty and M Sur, MeCP2-regulated miRNAs control early human neurogenesis through differential effects on ERK and AKT signaling, Molecular Psychiatry, (2017).
- Cláudia C. Miranda, Tiago G. Fernandes, M. Margarida Diogo and Joaquim M.S. Cabral, Scaling up a chemically‐defined aggregate‐based suspension culture system for neural commitment of human pluripotent stem cells, Biotechnology Journal, 11, 12, (1628-1638), (2016).
- A G Chiocchetti, D Haslinger, J L Stein, L de la Torre-Ubieta, E Cocchi, T Rothämel, S Lindlar, R Waltes, S Fulda, D H Geschwind and C M Freitag, Transcriptomic signatures of neuronal differentiation and their association with risk genes for autism spectrum and related neuropsychiatric disorders, Translational Psychiatry, 6, 8, (e864), (2016).
- Joaquim M. S. Cabral and Sean P. Palecek, Editorial: Stem Cell Engineering, Biotechnology Journal, 10, 10, (1509-1510), (2015).
- André L. Rodrigues, Carlos A. V. Rodrigues, Ana R. Gomes, Sara F. Vieira, Sara M. Badenes, Maria M. Diogo and Joaquim M.S. Cabral, Dissolvable Microcarriers Allow Scalable Expansion And Harvesting Of Human Induced Pluripotent Stem Cells Under Xeno‐Free Conditions, Biotechnology Journal, 1800461, (2018).
- Serena Barral and Manju A. Kurian, Utility of Induced Pluripotent Stem Cells for the Study and Treatment of Genetic Diseases: Focus on Childhood Neurological Disorders, Frontiers in Molecular Neuroscience, 10.3389/fnmol.2016.00078, 9, (2016).
