Establishment and characteristics of white ear lobe chicken embryo fibroblast line and expression of six fluorescent proteins in the cells.
A white ear lobe chicken embryo (WELCE) fibroblast cell bank, containing 322 tubes of frozen cells, was successfully established from primary explants of 57 embryo samples. The cells were morphologically consistent with fibroblasts, and the growth curve was sigmoidal with a population doubling time (PDT) of 48 h. Karyotyping and G-banding indicated a total chromosome number of 2n=78; the rate of diploidy in the cell bank was 97.62%. The cells were also free from bacterial, fungal, viral and mycoplasma contamination. Analysis of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) isoenzymes ruled out cross-contamination between cells. In order to study exogenous gene expression, six fluorescent proteins were transfected into the WELCE cells. The transfection efficiency of these genes was between 10.1 and 41.9%. The corresponding fluorescence was distributed throughout the cytoplasm and nucleus 24h after transfection. The results indicate that the quality of the cell line meet the quality requirements of the ATCC (American Type Culture Collection).
Wu
Hongmei
H
Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100094, PR China.
Guan
Weijun
W
Li
Han
H
Ma
Yuehui
Y
D000818:Animals; D002451:Cell Compartmentation / Q000502:physiology; D018929:Cell Culture Techniques; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D049109:Cell Proliferation; D048430:Cell Shape / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D002642:Chick Embryo; D002875:Chromosomes / Q000235:genetics*; D003593:Cytoplasm / Q000378:metabolism / Q000648:ultrastructure; D004171:Diploidy; D004734:Energy Metabolism / Q000502:physiology; D004866:Equipment Contamination; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression / Q000235:genetics*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics*; D008291:Malate Dehydrogenase / Q000378:metabolism; D013997:Time Factors
D008164:Luminescent Proteins; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase