Establishment and Identification of Immortalized Skin Fibroblast Cell Line of Chinese Tree Shrew

PurposeTo provide a new experimental material to explore the role of skin fibroblasts and their extracellular matrix in dermis, the immortalized skin fibroblast line of Chinese tree shrew was established.

MethodsThe skin fibroblasts were separated and purified by digestion with trypsin from leg ministry skin of Chinese tree shrew. The skin fibroblasts were immortalized by simian virus 40 large T antigen (SV40T) transduction, and the immortalized cells were identified by morphological observation, immunofluorescence and karyotype identification after picked monoclonal subculture and propagated to more than 50 generations of cells.

ResultsThe immortalized cells showed a similar morphology to that of the Chinese tree shrew primary skin fibroblasts with unaltered form, spindle and polygonal fibrous. The 58th generation cells were still in good condition. The growth curve showed immortalized cells grew vigorously. The cells were in the logarithmic growth phase on the 2nd or 3rd day, and entered the plateau phase on the 5th day. High fluorescence representing protein of tree shrew Vimentin and SV40T were detected by immunofluorescence. Karyotype analysis showed that the number of chromosomes was consistent with that of Chinese tree shrew, which means the cells obtained as immortalized tree shrew skin fibroblasts.

ConclusionA tree shrew skin fibroblasts cell line was successfully established, which could provide an experimental material for the study of damages and repairing about skin and other skin related diseases.