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Journal of Animal and Veterinary Advances
Year: 2012  |  Volume: 11  |  Issue: 23  |  Page No.: 4374 - 4381

Establishment and Biological Characteristics of Qing Kedan Chicken Embryonic Fibroblast Line for Genetic Resource Conservation

Hui Wang, Xiangchen Li, Changli Li, Wenxiu Zhang, Weijun Guan and Yuehui Ma    

Abstract: A fibroblast cell line of Qing Kedan chicken was successfully isolated, purified and cryopreseved through direct culturing of explants, serial passage and cryogenic techniques. Assays for biological characteristics were conducted and the results suggested that: the cells cultured with a Population Doubling Time (PDT) of about 20 h, constituted a typical fibroblast cell line in morphology; tests for microbial contamination regarding bacteria, fungi, viruses and mycoplasmas were exclusively negative; Lactate Dehydrogenase (LDH) and Malate Dehydrogenase (MDH) polymorphism analysis disproved the existence of cross-contamination from other cell lines; diploid cells making up 85-93% of the population verified the hereditary stability of the fibroblast line. pEGFP-N3, pEYFP-N1 and pDsRed1-N1 were transfected into the Qing Kedan chicken embryonic fibroblasts with transfection efficiencies between 12.6 and 39.7%. To locate their cellular distribution, transfected cells were observed at 24, 48 and 72 h with confocal microscopy and the fluorescences could be observed throughout the cytoplasm and nuclei of positive cells except in cryptomere vesicles. This research preserved the precious germplasm resource of Qing Kedan chicken at cell level and hence served as an enlightening reference for those of other poultry in the world.

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