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Brain Research Protocols
Volume 15, Issue 1, May 2005, Pages 52-58
 
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doi:10.1016/j.brainresprot.2005.03.004    How to Cite or Link Using DOI (Opens New Window)
Copyright © 2005 Elsevier B.V. All rights reserved.

Protocol

A modified method for generation of neural precursor cells from cultured mouse embryonic stem cells

Haiwei Xua, 1, Xiaotang Fanb, Corresponding Author Contact Information, 1, E-mail The Corresponding Author, Jun Tanga, Guangji Zhoua, Li Yanga, Xuan Wua, 2, Shingyong Liuc, 2, Jifu Qud, 2 and Hui Yangc, Corresponding Author Contact Information, E-mail The Corresponding Author

aDepartment of Physiology, The Third Military Medical University, Chongqing 400038, P.R. China bDepartment of Neurobiology, The Third Military Medical University, Chongqing 400038, P.R. China cDepartment of Neurosurgery, Xinqiao Hospital, The Third Military Medical University, Chongqing 400038, PR China dDepartment of Emergency, Southwest Hospital, The Third Military Medical University, Chongqing 400038, P.R. China

Accepted 23 March 2005. 
Available online 25 April 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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