A new method for isolation and culture of fibroblast-like synovial cells from adjuvant arthritis rabbits / 第二军医大学学报

ABSTRACT

[Abstract] Objective To establish a new, simple method for isolation and culture of fibroblast-like synoviocytes (FLSs) from adjuvant arthritis (AA) rabbits using suspended explant culture method. Methods Sixteen healthy New Zealand white rabbits, weighing 2.0-2.5 kg, were randomly divided into two groups the normal control group (n＝8) and the model group (n＝8). Rabbit models of AA were prepared by injection with complete Freund’s adjuvant at multiple sites and time points. The joint synovial layers were obtained. The FLSs of rabbits with AA were cultured by suspended explant culture method and explant culture method. The growth and morphological characteristics of cells were observed, and the cell viability was measured by CCK-8 assay. The expression of vimentin was detected by immunocytochemistry. Results The tarsometatarsal joints and toes of rabbits in the model group were swollen more obviously than those in the normal control group. After the toes were dissected, obvious microvascular proliferation and inflammation were observed in the model group. The results of H-E staining showed that synoviocytes in the normal control group were arranged in a regular manner like beads, while those in the model group were arranged in a disordered manner, suggesting that the AA model was successfully prepared. The operating durations required for suspended explant culture method and explant culture method were about 25 min and 1 h, respectively. The morphology and viability were similar for the cells obtained by the two methods. The positive rates of vimentin in the suspended explant culture method and explant culture method group were 98.01% and 98.35%, respectively. Conclusion Isolation and culture of FLSs by suspended explant culture method has been successfully established, and the method is simple and needs a short time.