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Abstract
The ability to differentiate mouse embryonic stem cells (ESC) to neural progenitors allows the study of the mechanisms controlling neural specification as well as the generation of mature neural cell types for further study. In this protocol we describe a method for the differentiation of ESC to neural progenitors using serum-free, monolayer culture. The method is scalable, efficient and results in production of ~70% neural progenitor cells within 4-6 days. It can be applied to ESC from various strains grown under a variety of conditions. Neural progenitors can be allowed to differentiate further into functional neurons and glia or analyzed by microscopy, flow cytometry or molecular techniques. The differentiation process is amenable to time-lapse microscopy and can be combined with the use of reporter lines to monitor the neural specification process. We provide detailed instructions on media preparation and cell density optimization to allow the process to be applied to most ESC lines and a variety of cell culture vessels.
Original language | English |
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Article number | e52823 |
Journal | JoVE: Journal of Visualized Experiments |
Issue number | 99 |
DOIs | |
Publication status | Published - 14 May 2015 |
Keywords
- Embryonic stem cells
- differentiation
- screening
- cell culture
- N2B27
- in vitro
- monolayer
- neural specification
- developmental biology
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Dive into the research topics of 'Neural differentiation of mouse embryonic stem cells in serum-free monolayer culture'. Together they form a unique fingerprint.Projects
- 1 Finished
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Strategic Award: Wellcome Trust Technology Platform
Blow, J. (Investigator), Lamond, A. (Investigator) & Owen-Hughes, T. (Investigator)
1/01/13 → 30/09/18
Project: Research
Student theses
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The regulation of mouse embryonic stem cell differentiation by Nrf2
Wongpaiboonwattana, W. (Author), Stavridis, M. (Supervisor) & Dinkova-Kostova, A. (Supervisor), 2017Student thesis: Doctoral Thesis › Doctor of Philosophy
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