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背景:利用骨髓间质干细胞(mesenchymalstemcells,MSCs)的多向分化性,选择合适的生物材料和适当的生长因子联合应用,可达到修复组织缺损的目的,MSCs细胞具有取材方便,对身体健康损害小,来源于自体干细胞诱导构建的组织不存在MHC限制等优点,所以日益受到人们的重视。目的:探讨人骨髓间充质干细胞向成骨和成软骨细胞诱导分化的能力。设计:单一样本研究。单位:苏州大学附属儿童医院骨科,苏州大学生命科学院生物技术研究所。材料:RPMI1640完全培养基,地塞米松,β-甘油磷酸钠,抗坏血酸,鼠抗人一抗,羊抗鼠二抗,链球菌亲和素(三抗),DAB(1:50),100mL/L胎牛血清,维生素C,转化生长因子-β1(TGF-β1)等。方法:选用不同代次的MSCs细胞,分别使用成骨和成软骨条件培养基培养,观察细胞的形态学变化,同时采用细胞化学和免疫组化的方法检测碱性磷酸酶,钙盐沉积,糖胺多糖分泌和I,Ⅱ胶原的表达。主要观察指标:骨髓基质干细胞向成骨细胞分化;骨髓基质干细胞向成软骨细胞分化。结果:MSCs细胞在地塞米松、维生素C和β-甘油磷酸钠的作用下,15d后可见细胞变为多边形,ALP和I型胶原染色阳性,形成钙结节,表现成骨细胞分化特点;而在维生素C和TGF-β1的作用下,7d可见细胞多为圆形,糖胺多糖和II型胶原染色阳性,表现成软骨细胞分化特点。
BACKGROUND: Utilizing the multi-directional differentiation of mesenchymal stem cells (MSCs) and selecting appropriate biomaterials and appropriate growth factors for the purpose of repairing tissue defects, MSCs have the advantages of convenient drawing and less damage to health , Derived from stem cells induced self-organized tissue does not exist MHC limits, etc., so more and more people’s attention. Objective: To investigate the ability of human bone marrow mesenchymal stem cells to differentiate into osteogenic and chondrogenic cells. Design: Single Sample Study. SETTING: Department of Orthopedics, Children’s Hospital Affiliated to Soochow University, Institute of Biotechnology, Suzhou University. Materials: RPMI1640 complete medium, dexamethasone, sodium β-glycerophosphate, ascorbic acid, mouse anti-human primary antibody, goat anti-mouse secondary antibody, streptavidin L fetal bovine serum, vitamin C, transforming growth factor-β1 (TGF-β1) and so on. Methods: Different generations of MSCs were selected and cultured in osteogenic and chondrogenic conditioned medium to observe the morphological changes of cells. Cytochemical and immunohistochemical methods were used to detect alkaline phosphatase, calcium deposition, Amidopolysaccharide secretion and collagen I and II expression. MAIN OUTCOME MEASURES: Bone marrow stromal stem cells differentiate into osteoblasts; Bone marrow stromal stem cells differentiate into chondrogenic cells. Results: MSCs cells became polygons after 15 days under the action of dexamethasone, vitamin C and β-glycerophosphate. Positive ALP and type I collagen staining formed calcium nodules, which showed the differentiation characteristics of osteoblasts; Under the action of vitamin C and TGF-β1, cells were mostly round after 7 days, and glycosaminoglycan and type II collagen were positive staining, which showed chondrocyte differentiation.