The aim of this work was to seek a different approach to conserving important domestic animals in imminent danger. A Qingyuan partridge chicken embryonic fibroblast line, containing 336 cryovials with 8x10(6) cells each, was successfully established from 60 Qingyuan partridge chicken embryos using explant culture and cryopreservation techniques. The cells were morphologically consistent with fibroblasts. The growth curve was a typical "S" shape having a detention phase, a logarithmic phase, and a plateau phase. The population doubling time was approximately 72 h. Tests for bacteria, fungi, viruses, and Mycoplasma were all negative. Isoenzyme polymorphism indicated that the genetic characteristics of the cell line were stable in vitro. Karyotyping analysis suggested that the chromosome number of a normal cell was 2n=78 and 93.4% of the entire population was diploid. The transfection efficiencies of 6 fluorescent proteins (pEGFP-C1, pEGFP-N3, pDsRed-N1, pEYFP-N1, pECFP-N1, and pECFP-mito) optimal at 48 h were from 15.1 to 39.8%. The cell line met all of the criteria from the American Type Culture Collection. Not only has the germline of this important chicken breed been preserved at the cell level, but also valuable material has been provided for genome, postgenome, and somacloning research. Moreover, the establishment of this technical platform may provide both technical and theoretical support for storing the genetic resources of other animals and poultry at the cell level.