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  • 刘琴,陈芳,朱以良,张宜*.一种新的佐剂性关节炎兔成纤维样滑膜细胞分离培养法[J].第二军医大学学报,2019,40(7):793-797    [点击复制]
  • LIU Qin,CHEN Fang,ZHU Yi-liang,ZHANG Yi*.A new method for isolation and culture of fibroblast-like synovial cells from adjuvant arthritis rabbits[J].Acad J Sec Mil Med Univ,2019,40(7):793-797   [点击复制]
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一种新的佐剂性关节炎兔成纤维样滑膜细胞分离培养法
刘琴,陈芳,朱以良,张宜*
0
(解放军中部战区总医院医学实验科, 武汉 430070
*通信作者)
摘要:
目的 采用悬浮组织块法建立一种新的、操作简单的佐剂性关节炎新西兰大白兔模型成纤维样滑膜细胞体外分离培养法。方法 选择健康新西兰大白兔16只,随机分为2组:正常组和模型组,每组8只。对模型组多点多次注射弗氏完全佐剂制备佐剂性关节炎模型,取膝关节滑膜组织,分别采用悬浮组织块法、组织块贴壁法分离培养佐剂性关节炎兔成纤维样滑膜细胞,观察细胞生长状况及形态特征,用CCK-8法检测细胞活力,免疫荧光法鉴定波形蛋白的表达情况。结果 模型组兔足跗关节及足趾部较正常组明显肿胀,解剖兔足趾部后肉眼可见模型组伴有明显的微血管增生和炎症。H-E染色结果显示,正常组兔膝关节滑膜细胞呈串珠样规则排列,而模型组滑膜细胞排列紊乱,提示佐剂性关节炎新西兰大白兔模型制备成功。悬浮组织块法和组织块贴壁法操作所需时间分别约为25 min、1 h,分离所得细胞的形态、活性相似,波形蛋白阳性率分别为98.01%、98.35%。结论 悬浮组织块法可成功分离培养出佐剂性关节炎兔成纤维样滑膜细胞,操作简单,所需时间短。
关键词:  类风湿关节炎  悬浮组织块培养  组织块贴壁培养  成纤维样滑膜细胞  细胞培养技术
DOI:10.16781/j.0258-879x.2019.07.0793
投稿时间:2019-03-28修订日期:2019-06-26
基金项目:
A new method for isolation and culture of fibroblast-like synovial cells from adjuvant arthritis rabbits
LIU Qin,CHEN Fang,ZHU Yi-liang,ZHANG Yi*
(Department of Medical Experiment, General Hospital of Central Theater Command of PLA, Wuhan 430070, Hubei, China
*Corresponding author)
Abstract:
Objective To establish a new, simple method for isolation and culture of fibroblast-like synoviocytes (FLSs) from adjuvant arthritis (AA) rabbits using suspended explant culture method. Methods Sixteen healthy New Zealand white rabbits, weighing 2.0-2.5 kg, were randomly divided into two groups:the normal control group (n=8) and the model group (n=8). Rabbit models of AA were prepared by injection with complete Freund's adjuvant at multiple sites and time points. The joint synovial layers were obtained. The FLSs of rabbits with AA were cultured by suspended explant culture method and explant culture method. The growth and morphological characteristics of cells were observed, and the cell viability was measured by CCK-8 assay. The expression of vimentin was detected by immunocytochemistry. Results The tarsometatarsal joints and toes of rabbits in the model group were swollen more obviously than those in the normal control group. After the toes were dissected, obvious microvascular proliferation and inflammation were observed in the model group. The results of H-E staining showed that synoviocytes in the normal control group were arranged in a regular manner like beads, while those in the model group were arranged in a disordered manner, suggesting that the AA model was successfully prepared. The operating durations required for suspended explant culture method and explant culture method were about 25 min and 1 h, respectively. The morphology and viability were similar for the cells obtained by the two methods. The positive rates of vimentin in the suspended explant culture method and explant culture method group were 98.01% and 98.35%, respectively. Conclusion Isolation and culture of FLSs by suspended explant culture method has been successfully established, and the method is simple and needs a short time.
Key words:  rheumatoid arthritis  suspended explant culture  explant culture  fibroblast-like synoviocytes  cell culture technigiques