Establishment of Big Bone chicken fibroblast cell bank and study of its biological characteristics

A fibroblast bank of Big Bone chicken was established using tissue adherent culture method. This cell bank included 29 embryo samples, had stocks of 147 cryogenically-preserved vials each containing 2~3Å~106 cells, and met all the cell line quality control standards established by the American Type Culture Collection (ATCC). The cells cultured in vitro showed the typical morphology of fibroblasts. The growth curve assumed the “S” shape, and the cell population doubling time (PDT) was about 30h. Cell viability was 98.1% before cryopreservation and 96.6% after recovery. All tests for microbial contamination were negative. The isoenzyme pattern was of species specificity. The frequency of diploid cells was 91%. The transfection efficiency of three fluorescent protein genes fluctuated between 10.6% and 26.5%. These results showed that the cells cultured in vitro grew well and had stable genetic properties. This research thus does not only preserve the poultry genetic resources of Big Bone chicken at the cell level, but also opens new ways for preserving important genetic resources of endangered animals in the form of somatic cells.