本文選題：骨髓間充質(zhì)干細(xì)胞 + 慶大霉素�。� 參考：《中南大學(xué)》2012年碩士論文

【摘要】：目的本文研究兔骨髓間充質(zhì)干細(xì)胞(Bone marrow stem cells, BMSCs)在體外分離培養(yǎng)、誘導(dǎo)分化為成軟骨的能力,并初步觀察不同濃度(0μg/m、10μg/ml、50μg/ml100μg/ml200μg/ml)慶大霉素對(duì)其增殖、分化成軟骨的影響。 方法無(wú)菌條件下,從8周齡家兔取下一側(cè)股骨及脛腓骨,通過(guò)全血培養(yǎng)法分離骨髓間充質(zhì)干細(xì)胞,接種于含有胎牛血清的DMEM培養(yǎng)液中培養(yǎng)傳代。每日以倒置顯微鏡觀察細(xì)胞形態(tài)及生長(zhǎng)情況。取第三代骨髓間充質(zhì)干細(xì)胞分五組,包含一個(gè)對(duì)照組(培養(yǎng)液為兔骨髓間充質(zhì)干細(xì)胞完全培養(yǎng)基：DMEM.10%胎牛血清、100U/m1青霉素、100μg/ml鏈霉素,不含慶大霉素)和四個(gè)含慶大霉素濃度組(分別是10μg/ml、50μg/ml、100μg/ml、200μg/ml加兔骨髓間充質(zhì)干細(xì)胞完全培養(yǎng)基)。采用MTT比色法進(jìn)行增殖活性比較,Ⅱ型膠原免疫細(xì)胞化學(xué)染色和細(xì)胞甲苯胺藍(lán)染色檢測(cè)成軟骨分化情況。 結(jié)果1.體外培養(yǎng)的骨髓間充質(zhì)干細(xì)胞呈扁平的梭形成纖維樣細(xì)胞生長(zhǎng),且細(xì)胞形態(tài)均一,傳代穩(wěn)定。2.MTT實(shí)驗(yàn)顯示慶大霉素在100μg/ml、200μg/ml抑制細(xì)胞增殖,其余各濃度組對(duì)細(xì)胞增殖無(wú)明顯影響3.誘導(dǎo)后細(xì)胞甲苯胺藍(lán)染色顯示100μg/ml組及200μg/ml組細(xì)胞細(xì)胞染色較淺,大部分細(xì)胞失染,余各組陽(yáng)性表達(dá)：Ⅱ型膠原免疫組織化學(xué)染色顯示100μg/ml組及200μg/ml組細(xì)胞弱陽(yáng)性表達(dá),余各組陽(yáng)性表達(dá)。 結(jié)論1兔骨髓間充質(zhì)干細(xì)胞取材簡(jiǎn)單方便,培養(yǎng)傳代后增殖迅速,生長(zhǎng)穩(wěn)定。2兔骨髓間充質(zhì)干細(xì)胞能向軟骨細(xì)胞誘導(dǎo)分化。3慶大霉素在一定濃度(10μg/ml、50μg/ml)下對(duì)骨髓間充質(zhì)干細(xì)胞的增殖和向軟骨分化無(wú)明顯影響,但高于一定濃度(100μg/ml,200μg/ml)則抑制骨髓間充質(zhì)干細(xì)胞的增殖和軟骨分化能力。
[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，

本文編號(hào)：1982246