Isolation, Culture and Identification of Sheep Skin Fibroblasts
HU Xinyu1, YAO Yian1, YU Kexin1, HU Qingqing1, JIANG Junfang2, JIANG Yongqing2, SHI Guoqing3, WAN Pengcheng3, DAI Rong3, GUAN Feng1*
The isolation and culture of sheep fibroblasts can provide a basic research platform for the functional studies of wool development related genes and molecular breeding. But the existing methods of isolation and culture by adhesion and trypsin digestion required long time and had low efficiency. The double enzyme digestion method and differential adherent screening method have not been applied in the separation of sheep skin fibroblasts. The aim of this study was to improve the efficiency and purity of sheep skin fibroblasts by optimizing the isolation and culture procedure. The ear skin tissue samples were taken from newborn lambs, and the primary skin fibroblasts were digested by trypsin and type I collagenase. After 2-3 days of culture, the cells were sub-cultured and isolated until the obtained fibroblasts had a high purity. This procedure takes about 7 days. The results of immunofluorescence identification showed that the purity of sheep skin fibroblasts was 100%. In this experiment, the optimized double enzyme digestion method and differential adhesion were used to isolate and purify sheep skin fibroblasts, which improved the isolation and culture efficiency of fibroblasts, greatly shortened the culture period and improved the cell purity.
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