Copyright © 2005 Elsevier B.V. All rights reserved.
Protocol
A modified method for generation of neural precursor cells from cultured mouse embryonic stem cells
Accepted 23 March 2005.
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Abstract
The pluripotency and high proliferative capacity of embryonic stem (ES) cells make them an attractive source of different cell types for biomedical research and cell replacement therapies. It has been demonstrated that ES cells can be induced into neural precursor cells (NPCs) under conditions. NPCs can be expanded in large numbers for significant periods of time to provide a reliable source of cells for transplantation in neurodegenerative disorders and injury of the central nervous system. This study describes a modified method for generation of NPCs from cultured mouse ES cells.
Keywords: Embryonic stem cells; Neural precursor cells; Cell culture
Neuroscience classification codes: Development and regeneration, Cell differentiation and migration
Article Outline
- 1. Introduction
- 2. Type of research
- 3. Time required
- 4. Materials
- 4.1. Animals
- 4.1.1. Special equipment
- 4.1.2. Chemicals and reagents
- 4.2. Solutions
- 4.2.1. Water
- 4.2.2. Phosphate-buffered saline (PBS)
- 4.2.3. Dulbecco's modified Eagle's medium (DMEM, high glucose)
- 4.2.4. Fetal bovine serum (FBS)
- 4.2.5. Poly-l-ornithine (15 μg/ml)
- 4.2.6. 0.25 mg/ml trypsin
- 4.3. Statistics
- 5. Detailed procedure
- 5.1. MEF preparation
- 5.2. ES cell culture and EBs formation
- 5.3. Alkaline phosphatase activity detection of embryonic stem cell
- 5.4. Selection of NPCs
- 5.5. Expansion of NPCs
- 5.6. NPC detection
- 5.7. Differentiation of NPCs
- 6. Results
- 6.1. In vitro differentiation of mouse ES cells to NPCs
- 6.2. Expansion and differentiation of NPCs in vitro
- 7. Discussion
- 7.1. Troubleshooting
- Acknowledgements
- References







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