Analysis of axonal growth in organotypic neural cultures
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.
Figure 1
Posted 01 Mar, 2016
Analysis of axonal growth in organotypic neural cultures
Posted 01 Mar, 2016
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
© Research Square 2021