Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học General Psychiatry cung cấp cho các bạn kiến thức về ngành y đề tài: Cell culture and passaging alters gene expression pattern and proliferation rate in rheumatoid arthritis synovial fibroblasts. | Neumann et al. Arthritis Research Therapy 2010 12 R83 http content 12 3 R83 RESEARCH ARTICLE Open Access Cell culture and passaging alters gene expression pattern and proliferation rate in rheumatoid arthritis synovial fibroblasts Elena Neumann 1 Birgit Riepl2 Anette Knedla1 Stephanie Lefevre1 Ingo H Tarner1 Joachim Grifka3 Jurgen Steinmeyer4 Jurgen Scholmerich2 Steffen Gay5 and Ulf Muller-Ladner1 Abstract Introduction Rheumatoid arthritis synovial fibroblasts RASF are key players in synovial pathophysiology and are therefore examined extensively in various experimental approaches. We evaluated whether passaging during culture and freezing has effects on gene expression and cell proliferation. Methods RASF were passaged for up to 8 passages. RNA was isolated after each passage and cDNA arrays were performed to evaluate the RNA expression pattern during passaging. In addition doubling time of the cells was also measured. Results From passages 2-4 mRNA expression did not change significantly. Gene expression in RASF started to change in passages 5-6 with 7-10 differentially expressed genes. After passages 7-8 more than 10 of the genes were differentially expressed. The doubling rate was constant for up to 5 passages and decreased after passages 6-8. After freezing gene expression of the second passage is comparable to gene expression prior to freezing. Conclusions The results of this study show that experiments which examine gene expression of RASF and shall reflect or imitate an in vivo situation should be limited to early culture passages to avoid cell culture effects. It is not necessary to stop culturing SF after a few passages but to keep the problems of cell culture in mind to avoid false positive results. Especially when large-scale screening methods on mRNA level are used. Of note freezing does not affect gene expression substantially. Introduction Predominant features of rheumatoid arthritis RA are synovial hyperplasia synovial cell .
