Objective Isolation, culture, and identification of dopaminergic neurons from newborn tree shrew to establish a cellular model and study abnormal dopaminergic neurons. Methods B27 and Glutamax were added to culture medium for in vitro culture of dopaminergic neurons from tree shrew. The cultured cells were identified by immunofluorescence. Results Cells isolated from newborn tree shrew were identified as dopaminergic neurons. The rate of tyrosine hydroxylase (TH)?positive neurons was (88.49 ± 2.4)%. Conclusions This method can be used to culture dopaminergic neurons from tree shrew in vitro.