✍️ Author: Dr Eleni Christoforidou
🕒 Approximate reading time: 4 minutes
Neuronal culture, the practice of growing neurons outside the body, is an invaluable method in neuroscience research. By observing neurons in isolation, scientists can study their individual behaviours, physiological properties, and responses to various stimuli more effectively than in whole animals or human subjects.
Tissue Dissection and Dissociation: Brain or spinal cord tissue, often from embryonic or neonatal rodents, is dissected and dissociated into individual cells. The process involves a combination of mechanical trituration and enzymatic digestion.
Plating and Initial Culture: Cells are plated onto a substrate coated with materials like poly-D-lysine or laminin that promote adhesion. The cells are cultured in a suitable medium that supports neuronal survival and growth.
Maintenance: Neurons require careful maintenance, including regular medium changes and monitoring of their health and growth.
Neurons are post-mitotic and sensitive cells, which poses specific challenges for their culture:
Survival and Growth: Neurons have high metabolic demands and require specific growth factors and nutrients, often provided by glial cells in vivo. Co-culture with glial cells or supplementation with specific factors can support neuronal survival and growth in culture.
Neuronal Polarity: In vivo, neurons develop a highly polarised morphology with distinct axons and dendrites. Reproducing this polarity in culture can be challenging.
Connectivity: In vivo, neurons form complex networks of synaptic connections. In culture, neurons can form synapses, but reproducing the complex connectivity of the brain is not yet feasible.
Despite these challenges, neuronal cultures have proved instrumental in studying various aspects of neuroscience:
Neuronal Development: By observing neurons in culture, researchers can study processes like axon guidance, dendrite branching, and synapse formation in detail.
Neuronal Physiology: Neuronal cultures allow for detailed studies of neuronal electrophysiology, including action potential firing and synaptic transmission.
Disease Modelling: Neurons derived from patients or genetically engineered models can be used to study disease mechanisms and test potential therapeutics.
In the context of neurodegenerative diseases like ALS, neuronal cultures offer several advantages:
Disease Mechanisms: Culturing neurons expressing disease-associated mutations can help researchers understand how these mutations disrupt neuronal function.
Drug Testing: Potential therapeutics can be tested on cultured neurons to assess their effects on neuronal health and function.
Patient-Specific Models: With the advent of induced pluripotent stem cells (iPSCs), neurons can be derived from patients, allowing for the study of disease mechanisms in a patient-specific context.
Neuronal culture, while not without its challenges, is a powerful tool in neuroscience. It allows researchers to observe neurons in isolation, providing unique insights into neuronal development, function, and disease mechanisms. The continued refinement of neuronal culture techniques will undoubtedly contribute to our understanding of the nervous system and the development of therapeutics for neurodegenerative diseases like ALS.