The zebra finch Taeniopygia guttata is a model object of neurobiology ethology, cell and developmental biology. We describe a detailed protocol for successful establishment and maintaining of primary cell lines of the zebra fnch using enzymatic method. The cells obtained are of various morphology resembling keratinocytes, fibroblasts, and melanocytes. Zebra finch cell lines derived in this way can be cryopreserved and successfully recovered to provide a source for cytological and cytogenetic studies. The protocol can be used as a basis for cell culture establishment for any songbird.