Chicken Embryo Spinal Cord Slice Culture Protocol

Tubby, Kristina C.; Norval, Dee; Price, Stephen R.

doi:10.3791/50295-v

Cited by 1 publication

(2 citation statements)

References 10 publications

(11 reference statements)

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“…Here, we established a method combining in ovo electroporation and the culture of organotypic tissue slices to study exogenous gene functions in the CNS during chicken embryo development. Some of these methods have been established for the CNS tissue slice culture during chicken embryo development . Compared with the established methods, we have combined in ovo electroporation with tissue slice culture to study the function of exogenous genes in the CNS during chicken embryo development.…”

Section: Discussionmentioning

confidence: 99%

“…Today, organotypic brain slice culture is mainly applied to mouse and rat tissue, as well as for disease modelling, such as with transgenic mice . However, there are few reports related to the study of organotypic slice culture using chicken embryos . The chicken embryo is a good animal model, used for both developmental biology and neurobiology studies.…”

Section: Introductionmentioning

confidence: 99%

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Organotypic slice culture based on in ovo electroporation for chicken embryonic central nervous system

Yang

et al. 2018

J Cellular Molecular Medi

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Organotypic slice culture is a living cell research technique which blends features of both in vivo and in vitro techniques. While organotypic brain slice culture techniques have been well established in rodents, there are few reports on the study of organotypic slice culture, especially of the central nervous system (CNS), in chicken embryos. We established a combined in ovo electroporation and organotypic slice culture method to study exogenous genes functions in the CNS during chicken embryo development. We performed in ovo electroporation in the spinal cord or optic tectum prior to slice culture. When embryonic development reached a specific stage, green fluorescent protein (GFP)‐positive embryos were selected and fluorescent expression sites were cut under stereo fluorescence microscopy. Selected tissues were embedded in 4% agar. Tissues were sectioned on a vibratory microtome and 300 μm thick sections were mounted on a membrane of millicell cell culture insert. The insert was placed in a 30‐mm culture dish and 1 ml of slice culture media was added. We show that during serum‐free medium culture, the slice loses its original structure and propensity to be strictly regulated, which are the characteristics of the CNS. However, after adding serum, the histological structure of cultured‐tissue slices was able to be well maintained and neuronal axons were significantly longer than that those of serum‐free medium cultured‐tissue slices. As the structure of a complete single neuron can be observed from a slice culture, this is a suitable way of studying single neuronal dynamics. As such, we present an effective method to study axon formation and migration of single neurons in vitro.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%