Abstract: Taking Small Tail Han Sheeps ear margin as the study materials, we succeeded in establishing a fibroblast cell bank which content of samples was 52 by the method of explant culture directly and cryopreservation technique. The biological analysis results showed that population doubling time (PDT) of cells was approximately 24 h, diploid account for 90%～96% in chromosome. Confirmed by isoenzyme analysis,this cell line had no cross-contamination.The test results of the bacterium, fungi, virus, mycoplasma were negative. The transfection rate was high, and the extro-gene can be effectively expressed. Every indexes of the cell bank met all the standard quality controls of ATCC. It had not only preserved the germplasm resources of this important Small Tail Han Sheep on the cell level but also provided valuable material for the research of genome, postgenome and somacloning. Meanwhile, the setting-up of this technological platform will offer technology and theory for storing the cell genetic resources of other animals and poultry.