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Method Article
Analysis of axonal growth in organotypic neural cultures
Xavier Navarro
Neuroplasticity and Regeneration
Corresponding Author
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This work is licensed under a CC BY-NC 3.0 License. Read Full License
Organotypic cultures are multicellular in vitro models that preserve both cytoarchitecture and cell interactions that form the tissue, providing a closer approximation to in vivo models in comparison with dissociated cell cultures. Previous studies in our lab proposed a method of dorsal root ganglia (DRG) and spinal cord slice (SC) organotypic 3D cultures to study motor and sensory axonal regeneration. Although these models are useful to study how different factors and substrates affect axonal growth, manual sample analysis can be inaccurate, tiresome and high time-consuming. Therefore, we have developed a computer-aided method, using the Neurite-J plug-in, to analyze the neurite outgrowth in organotypic cultures, that can also be applied to other types of explants. This program, implemented as a plug-in for ImageJ software, markedly reduces the time needed in the manual analysis, improves the accuracy and increases the amount of information obtained from each sample. Therefore, these organotypic 3D cultures and the computed aired method are a powerful and useful tool to obtain valuable data of neurite growth in different conditions. The objective of the present work is to provide the protocol of our DRG and SC slice cultures, from the animal to the image analysis, that will allow studying neurite outgrowth in a reliable in vitro model.
See figure in Figures section.
Keywords
Axonal growth, Spinal cord slice culture, Dorsal root ganglia culture, Neurite-J, ImageJ
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Associated Publications
In vitro comparison of motor and sensory neuron outgrowth in a 3D collagen matrix
by Ilary Allodi, Mónica-Sofía Guzmán-Lenis, Joaquim Hernàndez, +2
Journal Of Neuroscience Methods (08 September, 2015)
Neurite-J: An Image-J plug-in for axonal growth analysis in organotypic cultures
by A. Torres-Espín, D. Santos, F. González-Pérez, +2
Journal Of Neuroscience Methods (08 September, 2015)
FGF-2 Low Molecular Weight Selectively Promotes Neuritogenesis of Motor Neurons In Vitro
by Ilary Allodi, Laura Casals-Díaz, Eva Santos-Nogueira, +3
Schwann cells transduced with a lentiviral vector encoding Fgf-2 promote motor neuron regeneration following sciatic nerve injury
by Ilary Allodi, Vasil Mecollari, Francisco González-Pérez, +5
NKCC1 Activation Is Required for Myelinated Sensory Neurons Regeneration through JNK-Dependent Pathway
by L. Modol, D. Santos, S. Cobianchi, +3
Journal Of Neuroscience (08 September, 2015)
Focal release of neurotrophic factors by biodegradable microspheres enhance motor and sensory axonal regeneration in vitro and in vivo
by Daniel Santos, Guido Giudetti, Silvestro Micera, +2
Substratum preferences of motor and sensory neurons in postnatal and adult rats
by Francisco Gonzalez-Perez, Albert Alé, Daniel Santos, +5
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This is a preprint. It has not completed peer review.
Method Article
Analysis of axonal growth in organotypic neural cultures
Xavier Navarro
Neuroplasticity and Regeneration
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 01 Mar, 2016
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Organotypic cultures are multicellular in vitro models that preserve both cytoarchitecture and cell interactions that form the tissue, providing a closer approximation to in vivo models in comparison with dissociated cell cultures. Previous studies in our lab proposed a method of dorsal root ganglia (DRG) and spinal cord slice (SC) organotypic 3D cultures to study motor and sensory axonal regeneration. Although these models are useful to study how different factors and substrates affect axonal growth, manual sample analysis can be inaccurate, tiresome and high time-consuming. Therefore, we have developed a computer-aided method, using the Neurite-J plug-in, to analyze the neurite outgrowth in organotypic cultures, that can also be applied to other types of explants. This program, implemented as a plug-in for ImageJ software, markedly reduces the time needed in the manual analysis, improves the accuracy and increases the amount of information obtained from each sample. Therefore, these organotypic 3D cultures and the computed aired method are a powerful and useful tool to obtain valuable data of neurite growth in different conditions. The objective of the present work is to provide the protocol of our DRG and SC slice cultures, from the animal to the image analysis, that will allow studying neurite outgrowth in a reliable in vitro model.
Figure 1
Associated Publications
In vitro comparison of motor and sensory neuron outgrowth in a 3D collagen matrix
by Ilary Allodi, Mónica-Sofía Guzmán-Lenis, Joaquim Hernàndez, +2
Journal Of Neuroscience Methods (08 September, 2015)
Neurite-J: An Image-J plug-in for axonal growth analysis in organotypic cultures
by A. Torres-Espín, D. Santos, F. González-Pérez, +2
Journal Of Neuroscience Methods (08 September, 2015)
FGF-2 Low Molecular Weight Selectively Promotes Neuritogenesis of Motor Neurons In Vitro
by Ilary Allodi, Laura Casals-Díaz, Eva Santos-Nogueira, +3
Schwann cells transduced with a lentiviral vector encoding Fgf-2 promote motor neuron regeneration following sciatic nerve injury
by Ilary Allodi, Vasil Mecollari, Francisco González-Pérez, +5
NKCC1 Activation Is Required for Myelinated Sensory Neurons Regeneration through JNK-Dependent Pathway
by L. Modol, D. Santos, S. Cobianchi, +3
Journal Of Neuroscience (08 September, 2015)
Focal release of neurotrophic factors by biodegradable microspheres enhance motor and sensory axonal regeneration in vitro and in vivo
by Daniel Santos, Guido Giudetti, Silvestro Micera, +2
Substratum preferences of motor and sensory neurons in postnatal and adult rats
by Francisco Gonzalez-Perez, Albert Alé, Daniel Santos, +5