Isolation of Infiltrating Leukocytes from the Spinal Cord of Mice
从小鼠脊髓分离浸润性白细胞
发布: 2013年05月20日第3卷第10期 DOI: 10.21769/BioProtoc.775 浏览次数: 12408
评审: Xuecai Ge
PDF 
Q&A
Abstract
The infiltration of leukocytes into the central nervous system (CNS) is a common feature of many neuroinflammatory diseases such as multiple sclerosis, and also occurs during certain microbial infections such as by West Nile Virus. Here, we describe a method to isolate leukocytes from the spinal cords of mice. This method can be used for the characterization of leukocyte populations that infiltrate the spinal cord, and to perform functional studies with the isolated cells. The CNS of naive mice is infiltrated by very low numbers of leukocytes, however, upon inflammation increased numbers of mononuclear cells traffic to the CNS. The number of leukocytes that can be isolated roughly correlates with the degree of inflammation.
Keywords: Experimental Autoimmune Encepahlomyelitis (实验性自身免疫性脑脊髓炎)
CD137 (CD137)
4-1BB (4-1BB)
Infiltration (浸润)
Spinal cord (脊髓)
Materials and Reagents
- 1x PBS and 10x PBS
- Surgical instruments: scissors, forceps, scalpel
- Percoll (Sigma-Aldrich, catalog number: P4937 )
Note: prepare isotonic Percoll solution stock (one part 10x PBS and 9 parts Percoll).
Equipment
- Centrifuge with swing-out bucket rotor (e.g. Eppendorf, model: 5810 R )
- 75 mm petri dish
- 18 G and 27 G needle (BD Biosciences)
- 10 ml syringe (BD Biosciences)
- 15 and 50 ml conical centrifuge tubes
- Plastic Pasteur pipette
- 70 μm cell strainer (BD Biosciences, catalog number: 352350 )
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Martinez Gomez, J. M., Gasser, S. and Schwarz, H. (2013). Isolation of Infiltrating Leukocytes from the Spinal Cord of Mice. Bio-protocol 3(10): e775. DOI: 10.21769/BioProtoc.775.
- Martinez Gomez, J. M., Croxford, J. L., Yeo, K. P., Angeli, V., Schwarz, H. and Gasser, S. (2012). Development of experimental autoimmune encephalomyelitis critically depends on CD137 ligand signaling. J Neurosci 32(50): 18246-18252.
Download Citation in RIS Format
分类
免疫学 > 免疫细胞分离
神经科学 > 神经系统疾病
您对这篇实验方法有问题吗？
在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。
提问指南
+ 问题描述
写下详细的问题描述，包括所有有助于他人回答您问题的信息（例如实验过程、条件和相关图像等）。
发布问题
