In Vitro Cell.Dev.Biol.—Animal (2011) 47:280–282 DOI 10.1007/s11626-011-9392-9 REPORT Zebra finch cell lines from naturally occurring tumors Yuichiro Itoh & Arthur P. Arnold Received: 7 November 2010 / Accepted: 26 January 2011 / Published online: 27 February 2011 / Editor: J. Denry Sato # The Author(s) 2011. This article is published with open access at Springerlink.com Abstract The zebra finch (Taeniopygia guttata) has been intensively studied in many research fields including neuroscience, behavioral neurobiology, and evolution of the genome. Although numerous molecular and genomic resources are available for this model species, immortalized cell lines have been lacking. We have established two zebra finch cell lines derived from spontaneous tumors. ZFTMA is a tetraploid female cell line and G266 as a diploid male cell line. These first zebra finch cell lines should facilitate development of research on this model species. Keywords Zebra finch . Cell line . Immortalized . Male . Female Introduction The zebra finch, an oscine songbird (order Passeriformes), has been extensively studied as a model for the behavioral neurobiology of learning, the role of adult neurogenesis, the neural basis of sensory perception and cognition, sexual differentiation of the brain, and evolution of the genome. The study of songbirds has contributed to numerous fields in neurobiology, neuroendocrinology, and evolution. (Ziegler and Marler 2004). The zebra finch was the second avian genome to be sequenced (Warren et al. 2010). Although numerous genomic resources are now available for this important animal model, there have been no available Y. Itoh (*) : A. P. Arnold Department of Integrative Biology and Physiology, University of California, UCLA 610 Charles E. Young Drive South, Room 1146, Los Angeles, CA 90095-1606, USA e-mail: yitoh@ucla.edu immortalized cell lines for intensive molecular and cell biology research including transfection of specific genes or recombinant protein expression. To establish a valuable cell line resource, we isolated cells from naturally occurring tumorous tissues. Although such tumors are rare, over the years, we found two birds with tumor-like abnormal tissue in our zebra finch colony in Los Angeles. One female bird (Brown 264), which we established ZFTMA cell line from, had a left thigh that was almost 20–30 times larger than the right one. This bird had complete female plumage and normal ovary, but its liver was almost ten times larger and kidney size was also about six times larger than normal. Another bird (Gray 266), which we established G266 cell line from, had a tumor-like tissue under the skin of his forehead. This bird had male plumage and normal testis, and the other parts of his body including the internal organs did not have any abnormality. Materials and Methods In both cases, the bird was anesthetized and sacrificed, and tumor was dissected out. The tissue was washed with phosphate-buffered saline, minced, and transferred into 0.5% collagenese B (Roche, Indianapolis, IN)/Hank’s balanced salt solution. The tissue was incubated at 37°C for 1–3 h and mechanically dissociated. Ten milliliters of Dulbecco’s Modified Eagle Medium (DMEM; Sigma, St. Louis, MO) supplemented with 36.7 mmol/L glucose, 10% v/v heat-inactivated fetal bovine serum (Gibco BRL, Gaithersburg MD), 2% v/v heat-inactivated chicken serum (Gibco BRL), and antibiotics (100 U/ml penicillin G, 100 μg/ml streptomycin sulfate) were added to the cells, and it was further dissociated by pipetting up and down. The