Culture, Purification And Identification Of Synovial Fibroblasts From Rats With Type Ii Collagen-Induced Arthritis
Keywords
Synovial Fibroblasts, Collagen-Induced Arthritis Rat Model, Primary Culture, Rheumatoid Arthritis
Abstract
Fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA) are the most ideal cell source for studying the pathogenesis of RA and screening RA drugs. However, for many non-medical institutions of universities and colleges For scientific research institutes, since the synovial tissue of RA patients is not easy to obtain, it is difficult to perform operations at the cellular level. Therefore, a collagen-induced arthritis (CIA) rat model was established, and the synovial tissue of the knee joint was successfully isolated and Primary culture and purification of synovial fibroblasts will effectively solve the problem of the source of RA FLS and lay the foundation for subsequent experiments. In this experiment, Wistar rats were induced to suffer from arthritis by intradermal and subcutaneous injections of chicken type II collagen twice. The CIA model was established through the arthritis index score and HE staining of the knee joint; the synovial tissue of the knee joint was isolated, and the cells were isolated using type II collagenase and EDTA-free trypsin digestion and subcultured. When they were passed to the third generation, the cells Immunofluorescence was used to detect the VCAM-1 protein content in the cells to identify the purity of the cells. The results showed that a large number of rat synovial fibroblasts were obtained after digestion and culture of the synovial tissue; the cells observed under the microscope were spindle-shaped and dendritic, with Characteristics of fibroblast-like cells; VCAM-1 protein-positive cells reached more than 98%, confirming that the isolated cells were synovial fibroblast-like cells with extremely high purity. This study established a relatively simple and rapid method for obtaining a large number of synovial fibroblasts. This method provides a wealth of experimental materials for research on the pathogenesis of RA and drug prevention and treatment.
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Original research was done by Qiu Haiwen, Liu Mei, Liu Zhenzhou, Sun Shengnan, Li Hui, Ma Xuemei
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