Objective To establish an easier and better method for primary culture of rabbit fibroblast-like synoviocytes(FLS) by modified tissue cell culture. Method Healthy rabbit joint synovial layers were cut into small pieces and siphoned off water attached on the tissue with sterile filter paper and then cultured. The growth status and morphology were observed. The growth curve of the 3^rd passage cells was measured by CCK-8 assay, and the expression of vimentin was detected by immunocytochemistry. Results The FLS cultured in vitro exhibited a spindle-shaped appearance and could rapidly expand. The cell growth curve was typical of S type, and the cells highly expressed vimentin. Conclusions Primary culture of rabbit FLS by the improved explant culture method is successfully established. The method is simple and highly efficient. It provideds a new method for isolation of FLS.