Huang Xian-Zhang, Wang Qian, Zheng Lei, Chen Xiao, Xiao Ping, Xiong Shi-Long, Bao Jie, Ding Hai-Ming, Huang Wu-Jiao, Zhuang Jun-Hua
Laboratory Medicine Center, Nangfang Hospital, Southern Medical University, Guangzhou, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2009 Mar;29(3):462-5.
To isolate and characterize human rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs).
The synovial membrane tissues were obtained from 4 RA patients, 1 chondroma patient and 1 healthy subject and FLS were isolated by means of tissue culture. The cell morphology was observed by phase-contrast microscope and the cell surface markers were detected by flow cytometry.
The FLSs were successfully cultured from the synovial membrane tissues with good cell homogeneity after the third passage. The FLSs of the 3rd to 7th passages were stable and proliferated actively, followed by slow proliferation and aging since the 8th passage. Flow cytometry showed that the 4th-passage FLSs from the RA patients contained 99.04% CD90(+) cells, 2.73% CD3(+) cells, 0.29% CD3(-)CD19(+) cells, 2.81% CD3(-)CD16(+)CD56(+) cells, 5.89% CD14(+) cells, and 54.17% CD55(+) cells. The presence of interleukin-1 receptor type I (IL-1RI, 158.63-/+20.32 pg/ml) and IL-1beta (4.67-/+0.82 pg/ml) were detected in the cell culture supernatant of the 4th-passage FLSs from the RA patients by enzyme-linked immunosorbent assay ELISA.
FLSs from RA patients can be effectively culture by means of tissue culture, and the cultured FLSs show high expressions of CD90, IL-1RI and IL-1beta.
分离并鉴定人类风湿关节炎（RA）成纤维样滑膜细胞（FLS）。
从4例RA患者、1例软骨瘤患者和1例健康受试者获取滑膜组织，通过组织培养分离FLS。用相差显微镜观察细胞形态，用流式细胞术检测细胞表面标志物。
滑膜组织成功培养出FLS，第三代后细胞同质性良好。第3至7代FLS稳定且增殖活跃，自第8代起增殖缓慢并老化。流式细胞术显示，RA患者第4代FLS中CD90(+)细胞占99.04%，CD3(+)细胞占2.73%，CD3(-)CD19(+)细胞占0.29%，CD3(-)CD16(+)CD56(+)细胞占2.81%，CD14(+)细胞占5.89%，CD55(+)细胞占54.17%。通过酶联免疫吸附测定（ELISA）在RA患者第4代FLS的细胞培养上清液中检测到Ⅰ型白细胞介素-1受体（IL-1RI，158.63±20.32 pg/ml）和白细胞介素-1β（IL-1β，4.67±0.82 pg/ml）的存在。
通过组织培养可有效培养RA患者的FLS，培养出的FLS显示出CD90、IL-1RI和IL-1β的高表达。
