Development and optimization of a high-throughput 3D rat Purkinje neuron culture

Improved understanding of the mechanisms involved in neurodegenerative disease has been hampered by the lack of robust cellular models that faithfully replicate in vivo features. Here, we present a refined protocol for generating age-dependent, well-developed and synaptically active rat Purkinje neurons, responsive to paracrine factors and supporting a 3D cell network. Our model provides high experimental flexibility, high-throughput screening capabilities and reliability to elucidate Purkinje neuron function, communication and neurodegenerative mechanisms.