Isolation, culture, and characterization of duck primary neurons

The duck is a representative and good model for studying the development and physiological mechanisms of the nervous system (NS) in waterfowl. Neurons are the basic structural and functional units of NS, but there is no detailed method for cultured duck neurons in vitro. An efficient and simple method for duck neuron culture is reported in this study. First, the sfigpecific markers (NSE and GFAP, respectively) were used to explore the timing of the development of neurons and astrocytes during the duck embryonic stage (E5-E18). The cytomorphology of tissues and cells was tracked with the microscope at different time points. The brain tissues from 10-day-old duck embryos were determined as the optimal sampling embryo age for neuron culture. Then, the brain tissue isolation method (papain digestion) and cell suspension inoculation density (7 × 10⁵ cells/mL) were identified as the culture protocol to obtain target cells with high viability and high density. The purity of the cultured neurons was more than 95%. This experiment provides a supplement for the study of in vitro culture of waterfowl neurons and lays a good foundation for various subsequent studies.