A fibroblast line from embryos of Xiaoshan chicken was established successfully by direct culture of explants and cryopreservation techniques. The cells were morphologically consistent with fibroblasts, and the population doubling time (PDT) was about 46.72 h. According to karyotyping and G-banding, the diplontic cells with 78 chromosomes accounted for 98.58±1.27% of the total cells. The cells were tested for microbial contamination and they were free of infections from bacteria, fungi, viruses and mycoplasmas. There were no cross-contamination from other cell lines as revealed by isoenzyme polymorphism analysis of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH). Three fluorescent protein genes were transfected into the Xiaoshan chicken embryonic fibroblasts and the transfection efficiencies of these genes were between 12.72 and 35.89%. All the tests showed that the quality of the cell line conformed to the quality criteria of the American type culture collection (ATCC). This work had not only preserved the precious genetic resources of Xiaoshan chicken, but also explored a new protocol to preserve the endangered animal breeds.
Key words: Xiaoshan chicken, fibroblasts, genetic conservation, biological characteristics.
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