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滑膜干细胞向成骨细胞及软骨细胞分化的实验研究

发表时间:2014-12-15  浏览量:1430  下载量:571
全部作者: 曾建春,樊粤光,曾意荣,李杰,李飞龙
作者单位: 广州中医药大学第一附属医院三骨科
摘 要: 目的:探讨滑膜间充质干细胞(synovial mesenchymal stem cells,SMSCs)向成骨细胞及软骨细胞分化特性。方法:将第4代SMSCs以5×106个/cm2接种于3块6孔板,其中1块板诱导SMSCs向软骨细胞分化,剩下2块板诱导SMSCs向成骨细胞分化,每板4孔,2孔为空白对照孔,另2孔为观察组,2 d后换液,加入诱导液。空白组加入含10%胎牛血清(fetal bovine serum,FBS)的L-DMEM培养基继续培养,成骨诱导组在基础培养基中加入成骨诱导剂(含β甘油磷酸钠10 mmol/L,地塞米松0.1 μmol/L,维生素C 50 mg/L),软骨细胞组在基础培养基中加入软骨诱导剂[含转化生长因子β1(transforming growth factor-β1, TGF-β1)10 μg/L、地塞米松0.1 μmol/L、维生素C 50 mg/L]。在第14天取出1块6孔板做阿尔新蓝染色,1块6孔板做碱性磷酸酶染色;第20天取出剩余6孔板做茜素红染色。结果:SMSCs经软骨诱导液培养后,细胞体积增大,由梭形变为椭圆形状,部分细胞核偏位,呈肾形,培养14 d后可见细胞周围有基质分泌,阿尔新蓝染色呈阳性;SMSCs经成骨诱导液培养后,细胞从梭形变为类圆形,细胞体积明显增加,14 d时细胞集落的表面可见有颗粒状物质分泌,碱性磷酸酶染色呈阳性,21 d时茜素红染色呈阳性,表现出成骨细胞的特点。结论:SMSCs广泛存在于滑膜组织内,表现出干细胞特性,在特定条件下可向成骨细胞及软骨细胞分化。
关 键 词: 医学细胞生物学;滑膜间充质干细胞;成骨细胞;软骨细胞
Title: Experimental study on the differentiation of synovial mesenchymal stem cells into osteoblasts and chondrocytes
Author: ZENG Jianchun, FAN Yueguang, ZENG Yirong, LI Jie, LI Feilong
Organization: The Third Department of Orthopaedics, The First Affiliated Hospital of Guangzhou University of Chinese Medicine
Abstract: Objective: To explore the differentiation features of synovial mesenchymal stem cells (SMSCs) into the osteoblasts and chondrocytes. Methods: The fourth generation of SMSCs were seeded in 3 6-well plates by 5×106 cells/cm2, in one of the plates, SMSCs were induced to chondrocytes, in the remaining two boards, SMSCs were induced to differentiate into osteoblasts. Four holes in each plate, two holes were blank holes, and the other two holes were the observation group. 2 d later, the culture media were changed with inducing fluids. The control group was added L-DMEM medium containing 10% fetal bovine serum (FBS), the inducting group was added inducing agents on the basis of the L-DMEM medium [osteogenic group containing β glycerophosphate 10 mmol/L, dexamethasone 0.1 μmol/L, vitamin C 50 mg/L; the chondrogenic group containing transforming growth factor β1 (TGF-β1) 10 μg/L, dexamethasone 0.1 μmol/L, vitamin C 50 mg/L]. On the 14th day, a 6-well was removed for Alcian blue staining, a 6-well plate was done for alkaline phosphatase staining. On the 21st day, the remaining 6-well plate was done for Alizarin red staining. Results: Volume of SMSCs increased after induced by chondroinductive medium, the spindle changed into oval shape, some nuclei deviated and showed like kidney-shape. After 14 d, the surrounding cells matrix could be seen secreted by SMSCs, and Alcian blue staining was positive. After induced by osteogenic medium, SMSCs changed into oval from spindle in shape, volume increased significantly, on 14th day granular substance was visible on the surface of colony, alkaline phosphatase staining was positive, Alizarin red staining was also positive on 21st day, showing a characteristic of osteoblasts. Conclution: SMSCs were widely present in the synovial tissue, they showed the characteristics of stem cells which can be induced to differentiate into osteoblasts and chondrocytes under certain conditions.
Key words: medical cell biology; synovial mesenchymal stem cells; osteoblasts; chondrocytes
发表期数: 2014年12月第23期
引用格式: 曾建春,樊粤光,曾意荣,等. 滑膜干细胞向成骨细胞及软骨细胞分化的实验研究[J]. 中国科技论文在线精品论文,2014,7(23):2348-2354.
 
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