Equivalent osteoblastic differentiation function of human mesenchymal stem cells from rheumatoid arthritis in comparison with osteoarthritis
Equivalent osteoblastic differentiation function of human mesenchymal stem cells from rheumatoid arthritis in comparison with osteoarthritis
复制标题
DOI：
10.1093/rheumatology/kep044
复制
发表时间：
2009-06-01
期刊：
影响因子：
5.5
通讯作者：
Tomita, Tetsuya
中科院分区：
文献类型：
--
作者：
Morimoto, Daiki;Kuroda, Shoko;Tomita, Tetsuya
Objective.目的：探讨RA患者骨髓间充质干细胞（hMSCs）的成骨分化能力。肝素化骨髓穿刺液来自OA和RA患者。将单核细胞培养2周并进行集落形成测定。流式细胞仪分析细胞表型。第2代细胞在对照组中与β-甘油磷酸（bGP）一起培养，在分化组中与bGP、抗坏血酸和地塞米松一起培养。2周后进行ALP染色和活性测定。3周后，进行茜素红S测定。从培养2周和3周的细胞中提取总RNA。实时荧光定量PCR检测骨形成因子基因表达。OA和RA的细胞表型相同，内容物被认为是hMSCs。ALP活性和茜素红S测定结果显示，与对照组相比，OA和RA样品的分化组中的ALP活性水平较高。在OA和RA样品中，集落形成测定的结果是相同的。在OA和RA样本中，分化组的基因表达高于对照组。所有实验中OA和RA样本之间无显著性差异。骨关节炎和类风湿关节炎的hMSCs向成骨细胞分化的功能相似。
Objective. To evaluate the osteoblastic differentiation of human mesenchymal stem cells (hMSCs) in patients with RA.Methods. Heparinized bone marrow aspirate was obtained from patients with OA and RA. Mononuclear cells were cultured for 2 weeks and a colony-forming assay was performed. The phenotype of cells was analysed by flow cytometry. Passage 2 cells were cultured with -glycerophosphate (bGP) in the control group and bGP, ascorbic acid and dexamethasone in the differentiation group. After 2 weeks, ALP staining and activity were performed. After 3 weeks, Alizarin Red S assay was performed. Total RNA was extracted from cells cultured for 2 and 3 weeks. Gene expression of bone formation factor was examined by real-time PCR.Results. The phenotype of cells was identical in both OA and RA and the content was thought to be hMSCs. The results of ALP activity and Alizarin Red S assay showed higher levels in the differentiation group for both OA and RA samples compared with the control group. The results of a colony-forming assay were identical in both OA and RA samples. Gene expression in the differentiation group was higher than in the control group in both OA and RA samples. There was no significant difference between OA and RA samples in all experiments.Conclusion. The function of osteoblastic differentiation of hMSCs is similar between OA and RA.