动物造模,纤维样滑膜细胞炎症模型 z-bio 2022-11-02 253

　　Objective To establish the primary culture method of normal knee synovial cells in Sprague Dawley (SD) rats and the inflammation model of fibroblast like synovial cells induced by lipopolysaccharide.

　　Methods The synovium of 80~120g SPF young SD rats was isolated and cultured to the third generation. The FLS protein marker vimentin and proliferation function were detected by histochemical staining and EdU method. At the same time, synovial tissue was used as a control to detect the expression of characteristic proteins of FLS from the third to the eighth generation of primary culture, and to screen high purity FLS cells with physiological functions that can be used for subsequent experiments. After LPS induced FLS inflammation, detect IL-1 at different time points when LPS stimulated FLS β And TNF- α The time point of successful induction of FLS inflammation model by LPS was determined according to the experimental results. Finally, the expression of cytokines, proliferation function and characteristic proteins of FLS before and after LPS induction was detected to provide experimental data for the analysis of FLS inflammation model.

　　Results FLS primary cells were successfully cultured by 0.2% type I collagenase digestion. By detecting the protein marker vimentin, the purity of the third generation FLS was more than 98%. Through the detection of FLS characteristic proteins, the FLS cells with physiological functions that can be used for subsequent experiments were selected as the primary cells of the third to seventh generations. By analyzing the cytokines and characteristic proteins of LPS induced FLS, it is determined that 1 μ FLS cells were induced by g/mL LPS for 3 hours, and FLS inflammation model could be reproduced.

　　Conclusion The LPS induced FLS inflammatory model can be used as a cell model for the study of inflammatory arthritis in vitro.