Improve Research Reproducibility A Bio-protocol resource
Peer-reviewed
tooltip

Preparation of Primary Neurons from Rat Median Preoptic Nucleus (MnPO)

从大鼠正中视前核(MnPO)中制备原代神经元

Emmanuelle Berret Emmanuelle Berret

发布: 2013年12月05日第3卷第23期 DOI: 10.21769/BioProtoc.984 浏览次数: 10503

评审: Xuecai Ge

download pdf PDF PDF
ask Q&A
引用
收藏
Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of The Journal of Neuroscience, featuring study using the protocol.

Feb 2013

Advertisement

“科研可重复性与创新”系列网络研讨会
Special Issue on Protocols for Organoid Research
Technologies for RNA Detection
Presubmission Inquiry

实验方案合集

专注于特定技术或应用的、详细的、经过同行评审的实验方案合集

Cancer Research
Immunology
Developmental Biology
Microbiology
Neuroscience
Cell Imaging - A Special Collection for Cell Bio 2023
查看更多 more

相关实验方案

基于Langendorff灌流的小鼠心肌细胞简化分离方法

基于Langendorff灌流的小鼠心肌细胞简化分离方法

Mie S. Larsen [...] Tamzin Zawadzki

2026年07月20日 243 阅读

逆转录病毒制备后卫星细胞的分离培养与感染

逆转录病毒制备后卫星细胞的分离培养与感染

Chuanli Zhou [...] Elizabeth H. Chen

2026年07月20日 177 阅读

利用流式细胞术分离单核与双核肝细胞

利用流式细胞术分离单核与双核肝细胞

Yusuke V. Watanabe [...] Takeshi Katsuda

2026年07月20日 234 阅读

Abstract

Studying cell physiology is really important to understand their function and to determine action mechanisms taking place in these cells. Using brain slices can be sometime difficult to directly access to cells like neurons. Even if it more steps are needed to get cultured cells, this type of preparation allow a better access to neurons and provide a good way to study their basal properties. Here we describe a protocol of a short term primary neurons culture from MnPO, allowing to kept neurons in good condition to perform electrophysiological recordings.

Materials and Reagents

  1. Wistar rats (3 weeks old) (Charles River Laboratories International)
  2. Ketamine-xylasine mixture
  3. Pronase (Sigma-Aldrich, catalog number: 9036-06-0 )
  4. Thermolysin (Sigma-Aldrich, catalog number: 9073-78-3 )
  5. Bovine Serum Albumin (BSA) (Life Technologies, catalog number: 15561020 )
  6. Laminin (Sigma-Aldrich, catalog number: 114956-81-9 )
  7. Ruthenium red
  8. KCl (Sigma-Aldrich, catalogue number: 7447-40-7 )
  9. CaCl2 (Sigma-Aldrich, catalogue number: 10043-52-4 )
  10. MgCl2 (Sigma-Aldrich, catalogue number: 7786-30-3 )
  11. NaHCO3 (Sigma-Aldrich, catalogue number: 144-55-8 )
  12. NaH2PO4 (Sigma-Aldrich, catalogue number: 7558-80-7 )
  13. NaCl (Sigma-Aldrich, catalogue number: 7647-14-5 )
  14. Sucrose (Sigma-Aldrich, catalogue number: 57-50-1 )
  15. HEPES (Sigma-Aldrich, catalogue number: 7365-45-9 )
  16. D-Glucose (Sigma-Aldrich, catalogue number: 50-99-7 )
  17. Dissection solution (see Recipes)
  18. Artificial Cerebrospinal Fluid (aCSF) (see Recipes)

Equipment

  1. Bath
  2. Vibratome
  3. 0.45 μm sterilize filter
  4. 350 μm Sagittal slice containing MnPO
  5. Needle
  6. Pasteur pipettes
  7. Centrifuge
  8. 37 °C, 95% O2-5% CO2 Cell culture incubator
  9. 6 hole petri dish (Thermo Fisher Scientific, catalog number: 50-341-84 )
  10. Micro-cover glasses (Thermo Fisher Scientific, catalogue number: NC9216450 )
  11. 15 ml Falcon tubes
  12. Oxygen and Carbogen tank
  13. Binocular

Procedure

English
中文翻译

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:

  1. Berret, E. (2013). Preparation of Primary Neurons from Rat Median Preoptic Nucleus (MnPO). Bio-protocol 3(23): e984. DOI: 10.21769/BioProtoc.984.
  2. Berret, E., Nehme, B., Henry, M., Toth, K., Drolet, G. and Mouginot, D. (2013). Regulation of central Na+ detection requires the cooperative action of the NaX channel and α1 Isoform of Na+/K+-ATPase in the Na+-sensor neuronal population. J Neurosci 33(7): 3067-3078.

    3. ris ris Download Citation in RIS Format

分类

神经科学 > 细胞机理 > 细胞分离和培养

细胞生物学 > 细胞分离和培养 > 细胞分离

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。

post 发布问题
0 Q&A
pen 提交稿件