發(fā)布時間：2018-06-05 14:07

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

【摘要】：目的本文研究兔骨髓間充質(zhì)干細胞(Bone marrow stem cells, BMSCs)在體外分離培養(yǎng)、誘導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)慶大霉素對其增殖、分化成軟骨的影響。 方法無菌條件下,從8周齡家兔取下一側(cè)股骨及脛腓骨,通過全血培養(yǎng)法分離骨髓間充質(zhì)干細胞,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胞形態(tài)及生長情況。取第三代骨髓間充質(zhì)干細胞分五組,包含一個對照組(培養(yǎng)液為兔骨髓間充質(zhì)干細胞完全培養(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)干細胞完全培養(yǎng)基)。采用MTT比色法進行增殖活性比較,Ⅱ型膠原免疫細胞化學染色和細胞甲苯胺藍染色檢測成軟骨分化情況。 結(jié)果1.體外培養(yǎng)的骨髓間充質(zhì)干細胞呈扁平的梭形成纖維樣細胞生長,且細胞形態(tài)均一,傳代穩(wěn)定。2.MTT實驗顯示慶大霉素在100μg/ml、200μg/ml抑制細胞增殖,其余各濃度組對細胞增殖無明顯影響3.誘導后細胞甲苯胺藍染色顯示100μg/ml組及200μg/ml組細胞細胞染色較淺,大部分細胞失染,余各組陽性表達：Ⅱ型膠原免疫組織化學染色顯示100μg/ml組及200μg/ml組細胞弱陽性表達,余各組陽性表達。 結(jié)論1兔骨髓間充質(zhì)干細胞取材簡單方便,培養(yǎng)傳代后增殖迅速,生長穩(wěn)定。2兔骨髓間充質(zhì)干細胞能向軟骨細胞誘導分化。3慶大霉素在一定濃度(10μg/ml、50μg/ml)下對骨髓間充質(zhì)干細胞的增殖和向軟骨分化無明顯影響,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：1982246