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Isolation,Culture and Identification of Fibroblast-Like Synoviocytesfrom Collagen-Induced Arthritis(CIA)Rats(PDF)


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Isolation,Culture and Identification of Fibroblast-Like Synoviocytesfrom Collagen-Induced Arthritis(CIA)Rats
Qiu HaiwenLi HuiLiu ZhenzhouSun ShengnanMa XuemeiLiu Mei
School of Life Sciences,Nanjing Normal University,Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing 210023,China
synovial fibroblastscollagen-induced arthritis rat modelprimary culturerheumatoid arthritis
Fibroblast-like synoviocytes(FLS),which derived from rheumatoid arthritis(RA)patients,are the best cell source for the investigation of pathogenesis and treatment of RA. But it is difficult for some universities and institutions,to obtain human synovial tissue. So,it appears to be of essence to establish collagen-induced arthritis(CIA)rat model and there after obtain synovium for primary culture. The approach will be helpful to provide RA FLS efficiently,and lay the foundation for the follow-up experiment. Female Wistar rats were immunized intradermally with 1.5 mg native chicken collagen type II with an equal volume of Freund’s complete adjuvant. Seven days later,the rats were then given a subcutaneous booster injection of half the amount of CII emulsified in Freund’s incomplete adjuvant. When CIA model was established,the knee synovial tissue was separated and digested with type II collagenase and trypsin without EDTA. When the cells were cultured to the third generation,VCAM-1 protein in the cells was determined by immunostaining to identify cell purity. We obtained a large number of spindle-shaped cells. More than 98% of these cells were VCAM-1 positive,suggesting that these cells were rat synovial fibroblast-like cells in high purity. The establishment of a simple and rapid method for obtaining a large number of synovial fibroblasts will provides sufficient experimental materials for the pathogenesis and treatment of RA.


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Last Update: 2017-09-30