Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos

Sarah Powell; Amrit Vinod; Michele L. Lemons

doi:10.3791/51991

Aislamiento y cultivo de neuronas sensoriales disociadas De embriones de pollo

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Neuroscience

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JoVE Journal Neuroscience

Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos

Aislamiento y cultivo de neuronas sensoriales disociadas De embriones de pollo

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11:16 min

September 24, 2014

DOI: 10.3791/51991-v

Sarah Powell¹, Amrit Vinod¹, Michele L. Lemons¹

¹Department of Natural Sciences,Assumption College

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Overview

This article presents a method for isolating and culturing sensory neurons from chick embryos, providing a controlled environment for studying neuronal cell biology. The procedure is rapid, cost-effective, and reliable, allowing researchers to explore various aspects of neuronal function.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Developmental Biology

Background

Cell culture models are essential for studying neuronal biology.
Chick embryos serve as a valuable source for primary sensory neurons.
Isolation and culture techniques can enhance experimental control.
Immunocytochemistry is used to identify specific neuronal markers.

Purpose of Study

To develop a reliable method for culturing sensory neurons from chick embryos.
To provide detailed protocols for neuronal isolation and culture.
To facilitate the study of neuronal cell biology in a controlled environment.

Methods Used

Collection of dorsal root ganglia from embryonic chick embryos.
Dissociation of ganglia into a cell suspension.
Plating of dissociated cells on culture dishes.
Immunocytochemistry to identify neuronal markers such as NCA and beta one integrins.

Main Results

Successful isolation of primary sensory neurons from chick embryos.
Demonstration of neuronal presence through immunocytochemistry.
Establishment of a reliable culture method for sensory neurons.
Identification of growth cones and neurites in cultured neurons.

Conclusions

The method provides a robust platform for studying sensory neuron biology.
Immunocytochemistry confirms the successful culture of neurons.
This approach can be applied to various research questions in neuroscience.

Frequently Asked Questions

What are the main advantages of this method?

The method is rapid, inexpensive, and allows for detailed control over the culture environment.

What type of neurons are cultured using this technique?

Sensory neurons from chick embryos are isolated and cultured.

How are the neurons identified in culture?

Immunocytochemistry is used to identify specific neuronal markers.

What is the significance of using chick embryos?

Chick embryos provide a rich source of primary sensory neurons for research.

Can this method be adapted for other types of neurons?

While this method is specific to sensory neurons, similar techniques may be adapted for other neuronal types.

Modelos de cultivo celular proporcionan un control detallado de las condiciones ambientales y por lo tanto constituyen una plataforma poderosa para dilucidar numerosos aspectos de la biología de las células neuronales. Se describe un método rápido, barato y fiable para aislar, disociar, y la cultura neuronas sensoriales de los embriones de pollo. También se proporcionan detalles de la preparación de sustratos y inmunocitoquímica.

El objetivo general de este procedimiento es cultivar una población enriquecida de neuronas sensoriales primarias disociadas de embriones de pollo. Esto se logra recolectando ganglios de la raíz dorsal intactos del día embrionario, de siete a 10 embriones de pollo en un tubo cónico y posteriormente rompiéndolos en su suspensión celular disociada. Como segundo paso disociado, las células DRG se colocan en una placa de cultivo para su incubación.

Posteriormente, la placa de cultivo se enjuaga suavemente para desalojar preferentemente las neuronas. El paso final incluye la siembra de las neuronas sensoriales disociadas en los sustratos bien definidos, como cubreobjetos lavados con ácido y horneados precodificados con altas o bajas concentraciones de laminina. Se utiliza una inmunocitoquímica para mostrar la presencia de NCA e integrinas beta uno activadas en los cuerpos celulares, neuritas y conos de crecimiento de las neuronas sensoriales de pollo embrionario cultivadas disociadas.

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