Establishment and Biological Characteristics of Fibroblast Cell Lines in Newborn Kunming Dog

The objective of this study was to explore the method of isolation and cultivation of fibroblasts of Kunming dog in vitro.A newborn puppy ear skin of Kunming dog was used as the study material to establish fibroblast cell lines by the method of collagenase digestion and cryopreservation.The researches were designed to examine the biological characteristics of fibroblasts.The results showed that the cells were typical morphologic characteristics of fibrous and long spindle appearance.The average viability before freezing and after thawing was 93.3% and 90.8%,respectively.Additionally,the dynamic growth of the fibroblasts revealed a sigmoid curve with the population doubling time(PDT)of 32 h. Moreover,the karyotype result indicated the chromosome number of Kunming dog was 2n=78(XX)and diploid cells were dominant of 90%.In conclusion,the fetal fibroblast cell lines in Kunming dog were successfully established in vitro.It will not only preserve the national important genetic resources of Kunming dog at the cellular level but also provide valuable materials for somatic cell nuclear transfer of elite dog in the further.