Primary culture of hippocampal neurons of new-born rats / 基础医学与临床

ABSTRACT

Objective To establish a primary culture method of hippocampal neurons of new-born rats,and to observe the morphological characteristics at different developmental and differential stages.Methods The hippocampus was digested and the cells were seeded in a flask.The neurons were then transferred and re-seeded on cover glasses coated with poly-L-lysine.The neurons were identified by polyclonal antibody against neuron specific enolase(NSE).The morphology was observed under phase-contrast microscope.Results A large number of hippocampal neurons began to adhere to the cover glasses after 12～24 hours.They showed different shapes-shuttle,triangle,pyramidal,or nonregular after clinging to the plate.Their processes connected into nets and different in length and thickness.They were well developed on the 7th～10th day and survived as long as 28 days after seeding.Immunocytochemistry of NSE proved high purity.Conclusion The culture method of new-born rat hippocampal neurons in vitro is successful and can be used as an in vitro model of research.