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

J. Cell. Mol. Med., 2019 · DOI: 10.1111/jcmm.14080 · Published: March 1, 2019

Simple Explanation

This study introduces a method combining in ovo electroporation and organotypic slice culture to investigate gene functions within the developing central nervous system (CNS) of chicken embryos. In ovo electroporation allows for the introduction of exogenous genes into the spinal cord or optic tectum of the developing embryo. The tissue is then sliced and cultured, allowing for observation of neuronal behavior. This technique provides a new approach to study the dynamic changes of neurons under exogenous gene expression during chicken CNS development. The research also found that adding serum to the culture medium helped maintain the tissue structure and promoted the growth of neuronal axons. The method is particularly useful for studying single neuron dynamics.

Study Duration

7 days

Participants

Chicken embryos (stages 17-38) and E14.5 mice

Evidence Level

Not specified

Key Findings

1
The study successfully combined in ovo electroporation with organotypic slice culture for the chicken embryonic CNS, enabling the study of exogenous gene functions.
2
Adding serum to the culture medium (25% horse serum) significantly improved the maintenance of tissue morphology and promoted longer neuronal axon growth compared to serum-free medium.
3
The slice culture method allows for the observation of complete single neuron structures and their dynamic changes, making it suitable for studying neuronal migration and axon formation.

Research Summary

This study introduces and validates a method combining in ovo electroporation and organotypic slice culture to study gene function in the developing chicken central nervous system (CNS). The research demonstrates the importance of serum in the culture medium for maintaining tissue structure and promoting neuronal axon growth, with 25% horse serum showing significant benefits. The developed method is shown to be particularly useful for studying single neuron dynamics, including neuronal migration and axon formation, offering a valuable tool for neuroscience research.

Practical Implications

Enhanced Gene Function Studies

This method facilitates the study of exogenous gene functions in the developing chicken CNS, offering insights into neuronal development and related disorders.

Improved Culture Techniques

The finding that serum-containing medium enhances tissue structure and neuronal growth can inform and improve organotypic slice culture techniques for CNS research.

Single Neuron Dynamics Research

The method's suitability for observing single neuron dynamics opens new avenues for studying neuronal migration, axon formation, and the impact of genetic manipulations at the cellular level.

Study Limitations

1
The tissue slices lose their original morphological structure during culture, which may affect the physiological relevance of the findings.
2
The study acknowledges differences between in vitro slice culture conditions and in vivo conditions, particularly regarding GFAP and Iba1 expression.
3
Achieving successful transfection in the optic tectum in ovo electroporation is difficult and embryo survival rates are low.

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