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

Chick Neural Tube Explant Culture

鸡胚胎神经管外植体培养

Zhanna Alekseenko Zhanna Alekseenko
Elisabet Andersson Elisabet Andersson
José M. Dias José M. Dias

发布: 2015年10月05日第5卷第19期 DOI: 10.21769/BioProtoc.1608 浏览次数: 11180

评审: Oneil G. BhalalaPamela MaherGeoff Lau

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 Neuron, featuring study using the protocol.

Dec 2014

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

雏鸡绒毛尿囊膜（CAM）体内血管形成实验

雏鸡绒毛尿囊膜（CAM）体内血管形成实验

Zhenguo Chen [...] Xiaochun Bai

2013年09月20日 39112 阅读

用于高级功能分析的出生后啮齿动物全器官视网膜电转染方法

用于高级功能分析的出生后啮齿动物全器官视网膜电转染方法

Chien-Ting Huang [...] Chih-Tien Wang

2026年01月20日 397 阅读

RNA与蛋白质串联提取方法：最大化利用有限离体组织样本的实验平台

RNA与蛋白质串联提取方法：最大化利用有限离体组织样本的实验平台

Ciarán Kennedy [...] Monica de Gaetano

2026年03月20日 585 阅读

Abstract

The neural tube explant culture technique allows in vitro culturing of small pieces of neural tissue isolated from e.g., chick or mouse embryonic tissue in a matrix of collagen for defined periods of time. This method can be used to study the effects of defined molecules on developmental processes such as neural progenitor proliferation and neuronal differentiation and/or survival. Since the explant material can also be prepared from embryonic tissue electroporated with expression vectors, this technique can be adapted to study gene function in the presence of specific environmental signals. Different regions of the neural tube can also be isolated during the dissection step, allowing specific regions of the neural tube to be studied separately. Here, we present a neural tube explant culture method that we have used in several studies (Dias et al., 2014; Lek et al., 2010; Vallstedt et al., 2005).

Materials and Reagents

  1. 10 cm Petri dish (SARSTEDT AG & Co, catalog number: 82.1473 )
  2. Glass Pasteur pipette 150 mm (VWR International, catalog number: 612-1701 )
  3. Transfer pipette (SARSTEDT AG & Co, catalog number: 86.1174 )
  4. Tungsten wire (Goodfellow, catalog number: w005160 )
  5. 10 ml syringe (HARTMANN USA, Omnifix®, catalog number: 4617100V )
  6. Needle 20 G (Henke Sass Wolf, catalog number: 4710009040 )
  7. Fresh fertilized chick eggs (ideally from a local supplier)
  8. L-15 medium (Life Technologies, catalog number: 11415-049 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 11415-049”.
  9. Dispase I (Roche Diagnostics, catalog number: 04942086001 )
  10.  Heat inactivated Fetal bovine serum (FBS) ( Life Technologies, GibcoTM, catalog number: 10106-169 )
    Note: Currently, it is “Thermo Fisher Scientific, Gibco™, catalog number: 10106-169 ”.
  11. Dulbecco’s modified eagle’s medium 10x (MEM) (Sigma-Aldrich, catalog number: D2429 )
  12. Sodium hydroxide (NaOH) (EMD Millipore Corporation, catalog number: 1.06469.1000 )
  13. Collagen (3.1 mg/ml) (Pure Col) (Advanced BioMatrix, catalog number: 5005-B )
  14. Sodium bicarbonate 7.5% (BIC) (Life Technologies, catalog number: 25080 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 25080”.
  15. DMEM/F-12, GlutaMAXTM supplement (Life Technologies, catalog number: 31331-028 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 31331-028”.
  16. Neurobasal medium (Life Technologies, catalog number: 21103-049 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 21103-049”.
  17. Albumin from bovine serum (BSA) (Sigma-Aldrich, catalog number: A7906 )
  18. Glutamax (Life Technologies, catalog number: 35050-038 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 35050-038”.
  19. Penicillin Streptomycin (PEST) (Life Technologies, catalog number: 15140-122 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 15140-122”.
  20. N2-supplement (Life Technologies, catalog number: 17502-048 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 17502-048”.
  21. B27-supplement (Life Technologies, catalog number: 17504-044 )
    Note: Currently, it is “Thermo Fisher Scientific, GibcoTM, catalog number: 17504-044”.
  22. 2-mercaptoethanol (Sigma-Aldrich, catalog number: M3148 )
  23. Dispase I solution (see Recipes)
  24. Bedding solution (see Recipes)
  25. Growing media (see Recipes)

Equipment

  1. Humidified, 38 °C egg incubator (Grumbach, catalog number: 8012 )
  2. Humidified, 37 °C tissue culture incubator (Thermo Fisher Scientific, catalog number: 51030287 )
  3. 16 °C incubator (Gemini BV Laboratory，Liebherr, catalog number: 4294 F1-1 )
  4. Dissection microscope and light source (ZEISS, models: Zeiss Stemi SV11 and Zeiss KL1500LCD )
  5. Borosilicate glass capillaries (Harvard Apparatus, catalog number: GC100TF-10 )
  6. Nunc 4-well dishes for IVF (Thermo Fisher Scientific, catalog number: 144444 )
  7. Flaming/brown micropipette puller (SUTTER, catalog number: P-97 )
  8. Needle holder (Fine science tools, catalog number: 26016-12 )
  9. Tweezers (Fine science tools, catalog number: 11252-20 )
  10. Micro-scissors (Fine science tools, catalog number: 15000-00 )
  11. Dissection scissors (Fine science tools, catalog number: 14088-10 )
  12. Mouth aspirator system/Aspirator tube assemblies for calibrated microcapillary pipettes (Sigma-Aldrich, catalog number: A5177-5EA )

Procedure

English
中文翻译

文章信息

版权信息

© 2015 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Alekseenko, Z., Andersson, E. and Dias, J. M. (2015). Chick Neural Tube Explant Culture. Bio-protocol 5(19): e1608. DOI: 10.21769/BioProtoc.1608.

ris ris Download Citation in RIS Format

分类

神经科学 > 发育 > 外植体培养

细胞生物学 > 组织分析 > 组织分离

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