Development and Optimization of a Multilayer Rat Purkinje Neuron Culture

Cerebellum. 2024 Feb;23(1):101-111. doi: 10.1007/s12311-022-01510-4. Epub 2023 Jan 10.

Abstract

Elucidation of the mechanisms involved in neurodegenerative diseases of the cerebellum has been hampered by the lack of robust single cell models to study Purkinje neurons and replicate at the same time in vivo features. Cerebellar Purkinje neurons are difficult to grow in dispersed cell culture, and only limited work has been done using rat cells. We developed a refined protocol for growing rat Purkinje neurons from embryonic and postnatal tissue ex vivo that results in well-developed, mature, functional, and synaptically active neurons. The rat Purkinje neurons generated are responsive to paracrine factors and genetic manipulation, allowing great experimental flexibility at the single-cell level. This ex vivo model can be used to investigate disease mechanisms that disturb Purkinje neuron morphology, function, and communication in high- and low-throughput screening formats.

Keywords: Ex vivo culture system; Rat Purkinje neurons; Three-dimensional cerebellar network.

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cerebellum*
  • Neurons
  • Purkinje Cells* / physiology
  • Rats