不同浓度庆大霉素对兔骨髓间充质干细胞体外软骨形成能力影响的研究

发布时间：2018-06-05 14:07

  本文选题：骨髓间充质干细胞 + 庆大霉素　； 参考：《中南大学》2012年硕士论文

【摘要】：目的本文研究兔骨髓间充质干细胞(Bone marrow stem cells, BMSCs)在体外分离培养、诱导分化为成软骨的能力,并初步观察不同浓度(0μg/m、10μg/ml、50μg/ml100μg/ml200μg/ml)庆大霉素对其增殖、分化成软骨的影响。 方法无菌条件下,从8周龄家兔取下一侧股骨及胫腓骨,通过全血培养法分离骨髓间充质干细胞,接种于含有胎牛血清的DMEM培养液中培养传代。每日以倒置显微镜观察细胞形态及生长情况。取第三代骨髓间充质干细胞分五组,包含一个对照组(培养液为兔骨髓间充质干细胞完全培养基：DMEM.10%胎牛血清、100U/m1青霉素、100μg/ml链霉素,不含庆大霉素)和四个含庆大霉素浓度组(分别是10μg/ml、50μg/ml、100μg/ml、200μg/ml加兔骨髓间充质干细胞完全培养基)。采用MTT比色法进行增殖活性比较,Ⅱ型胶原免疫细胞化学染色和细胞甲苯胺蓝染色检测成软骨分化情况。 结果1.体外培养的骨髓间充质干细胞呈扁平的梭形成纤维样细胞生长,且细胞形态均一,传代稳定。2.MTT实验显示庆大霉素在100μg/ml、200μg/ml抑制细胞增殖,其余各浓度组对细胞增殖无明显影响3.诱导后细胞甲苯胺蓝染色显示100μg/ml组及200μg/ml组细胞细胞染色较浅,大部分细胞失染,余各组阳性表达：Ⅱ型胶原免疫组织化学染色显示100μg/ml组及200μg/ml组细胞弱阳性表达,余各组阳性表达。 结论1兔骨髓间充质干细胞取材简单方便,培养传代后增殖迅速,生长稳定。2兔骨髓间充质干细胞能向软骨细胞诱导分化。3庆大霉素在一定浓度(10μg/ml、50μg/ml)下对骨髓间充质干细胞的增殖和向软骨分化无明显影响,但高于一定浓度(100μg/ml,200μg/ml)则抑制骨髓间充质干细胞的增殖和软骨分化能力。
[Abstract]:Objective to study the ability of bone marrow stem cells, BMSCs) isolated from rabbit bone marrow mesenchymal stem cells to differentiate into cartilage in vitro, and to observe the effect of gentamicin on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells into cartilage. Methods Bone marrow mesenchymal stem cells (BMSCs) were isolated from the femur and tibia and fibula of 8 weeks old rabbits under aseptic condition. Bone marrow mesenchymal stem cells (BMSCs) were isolated by whole blood culture and cultured in DMEM medium containing fetal bovine serum. The morphology and growth of cells were observed by inverted microscope every day. The third generation of bone marrow mesenchymal stem cells (BMSCs) were divided into five groups, which were divided into five groups, including a control group (10% DMEM.10% fetal bovine serum) and 100 渭 g/ml streptomycin (100 渭 g/ml streptomycin) in rabbit bone marrow mesenchymal stem cells (BMSCs). 10 渭 g / ml ~ 50 渭 g / ml ~ (50 渭 g 路ml ~ (-1) 100 渭 g / ml ~ (200 渭 g/ml) and 10 渭 g / ml ~ (50 渭 g 路ml ~ (-1) 100 渭 g 路ml ~ (-1) 路L ~ (-1) of Gentamicin + rabbit bone marrow mesenchymal stem cell culture medium. The proliferative activity was compared by MTT colorimetry. The differentiation of chondrocytes was detected by type 鈪，

本文编号：1982246