Abstract: A Beijing fatty chicken embryo fibroblast cell line (NBLCHE2/2) was successfully established using the primary explant technique. Observations on morphology, dynamic growth and analysis of karyotype, isoenzymes of lactate dehydrogenase, malate dehydrogenase were carried out. The results showed that population doubling time of cells was 24 h; diploid cells were dominant of 76%～88%. The banding patterns of the isoenzymes of the two kinds of emzymes had significant difference between the Beijing fatty chicken embryo fibroblast cell line and other cell lines in our laboratory; tests for microbial conta mination of bacteria, fungi and yeasts, virus and mycoplasma were negative. The newly established cell line make the Beijing Fatty Chicken, a national important genetic resource preserved in cell level, as well as will provide an ideal experimental material for the genetic studies on the Beijing local chicken.