Umbilical cord mesenchymal stem cells (UMSCs) from fetal tissues have become a valuable source of cell regeneration therapy due to their low immunogenicity, high plasticity and antimicrobial peptides. Most researches about UMSCs mainly focused on humans, mice, rabbits and other mammals, but it is still unknown in broiler. To the best of our knowledge, we first established a broiler-derived UMSCs line from noncontroversial Wharton's jelly of umbilical cord. The detection of biological characteristics showed UMSCs showed the morphological characteristics of fibroblasts and could be cultured to 26 generations in vitro, and maintained diploid chromosomes (2n=78) by G-banding analysis. Cell cycle of UMSCs showed the rate of G0/G1 was about 76.7-80.9%. Moreover, UMSCs could express mesenchymal markers (CD29, CD90, CD71, and CD44), but not hematopoietic marker (CD34) and endothelial marker (CD31). Additionally, UMSCs could be efficiently induced to transdifferentiate into three germ layer cells, such as adipocytes, osteoblasts, chondrocytes, hepatocytes, epithelia, and neurogenic cells. These results demonstrated UMSCs have the essential characteristics of MSCs, and can be used as an attractive candidate for cell regenerative therapy and the study of regulatory mechanisms associated with selective differentiation.