Abstract: The Debao pony ear marginal tissue fibroblast cell line (NDPEM 2/2) was successfully established using collagenase technique. The characterizations of the cell line were identified as follows: the cells were adherent and of density limitation, and the population doubling time (PDT) of cells made with the collagenase technique was 35.9 h. Chromosome analysis showed that the frequency of cell chromosome number to be 2n = 64 was 91.3%～92.8%. Confirmed by isoenzyme analysis, this cell line had no cross-contamination. Tests for microbial contamination from bacteria, fungi, yeasts or mycoplasma were negative. This newly established cell line meets all the standard quality controls of ATCC. It will provide a precious genetic resource for the reservation of the Debao pony breed, as well as an effective experimental material for the genetic studies on Debao ponies. Collagenase technique was compared with primary explant technique in getting the primary cell at the same time, the former is very fast and the population doubling time (PDT ) of the later is 48 h.

Key words: Debao pony, Ear marginal tissue, Fibroblast cell line, Establishment, Identification