Human dopaminergic neuronal cells are isolated from fetal brain tissue obtained from agencies authorized to procure and distribute tissues for research. Dopaminergic neuronal cells are selected in tyrosine free medium supplemented with a mixture of growth factors. Actively growing population of cells are tested for tyrosine hydroxylase （TH） expression by immunocytochemistry. RFP-Expressing Human dopaminergic neuronal cells （RFP-HDaNCs） are selected by puromycin after HDaNCs are infected with lentiviruses expressing RFP. Cells are provided with more than 5x10[5] cells/ vial at passage 2. The cells are grown in regular tissue culture dishes and subcultured at a split ratio of 1 to 2 at confluence. We recommend not to use the cells beyond 4 passages. There may be a significant decrease in the number of tyrosine hydroxylase positive cells after 4 passages as they are not maintained in selective medium to allow the growth of only cells expressing tyrosine hydroxylase.
