Bai Chunyu

Establishment and characterization of a Chinese Game chicken embryo fibroblast bank

For preserving the genetic resources of the Chinese Game chicken and exploring a new method to preserve endangered animals, a Chinese Game chicken embryo fibroblast cell bank was established by the attachment culture method and the cryopreservation biotechniques. This bank includes 43 embryo samples and has stocks of 178 cryovials, with each containing 4.0×106 cells respectively. Quality assays and biological characteristics of the bank established showed that the cells cultured in vitro were all typical fibroblast and the cell population doubling time (PDT) was about 2 days. The cell viability before freezing was 97.37% ± 2.25% and 91.43% ± 3.22% after thawing. Tests for microbial contamination by bacteria, fungus, viruses and mycoplasmas were all negative. The frequency of cells having the diploid number of chromosomes (78) was 94.65% ± 3.29%. Isoenzyme analysis confirmed that there was no cross-contamitation in the culture. The fluorescence could be observed well-distributed in cytoplasm and nucleus except for some cryptomere vesicles at 24 h after transfection, and the expression efficiency of pEGFP-C1, pEGFP-N3, pEYFP-N1, pECFP-N1, pECFP-mito and pDsRed1-N1 were all between 9.5%∼34.6% at 24 h, 48 h and 72 h after transfection. This research provided both technical and theoretical support for preserving the genetic resources of other animals and poultry at the cell level.