發(fā)布時(shí)間：2018-11-11 15:47

【摘要】：目的研究骨髓間充質(zhì)干細(xì)胞(BMSCs)在體外向γ-氨基丁酸(GABA)能神經(jīng)元方向的分化。方法大鼠股骨骨髓分離出BMSCs,培養(yǎng)傳代,通過10%胎牛血清(FBS)、20 ng/ml表皮生長因子(EGF)和改良Eagle培養(yǎng)基(DMEM/F12)預(yù)誘導(dǎo)24 h,隨之加入含10 ng/ml堿性成纖維細(xì)胞生長因子(bFGF)、10 ng/ml骨形成蛋白-2(BMP-2)和10 ng/ml腦源性神經(jīng)營養(yǎng)因子(BDNF)作用48 h后,再以10μmol/L全反式維甲酸(ATRA)培養(yǎng)48 h,最后用40 mmol/L KCl刺激15 min完成誘導(dǎo)分化。對照組用10%FBS和DMEM/F12培養(yǎng)不添加任何誘導(dǎo)因子。形態(tài)學(xué)觀察和Western blot對分化后的細(xì)胞進(jìn)行鑒定分析。結(jié)果實(shí)驗(yàn)組細(xì)胞在形態(tài)學(xué)上表現(xiàn)出典型的神經(jīng)元樣細(xì)胞形態(tài),對照組無明顯變化;Western blot分析知實(shí)驗(yàn)組Ⅰ和實(shí)驗(yàn)組Ⅱ都有分化為GABA能神經(jīng)元的趨勢,但實(shí)驗(yàn)組Ⅱ的分化率高于實(shí)驗(yàn)組Ⅰ。結(jié)論BMSCs可在體外分化為GABA能神經(jīng)元。
[Abstract]:Objective To study the differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro to the neuronal orientation of 1-aminobutyric acid (GABA). Methods BMSCs were isolated from the bone marrow of the femur and then cultured for 24 hours by 10% fetal bovine serum (FBS), 20 ng/ ml epidermal growth factor (EGF) and modified Eagle's medium (DMEM/ F12). After 48 h, 10 ng/ ml of basic fibroblast growth factor (bFGF), 10 ng/ ml of bone morphogenetic protein-2 (BMP-2) and 10 ng/ ml of brain-derived neurotrophic factor (BDNF) were added, and then cultured for 48 h with 10. m u.mol/ L all-trans-retinoic acid (ATRA), and the induction of differentiation was completed by stimulation with 40 mmol/ L KCl for 15 min. The control group did not add any inducing factors with 10% FBS and DMEM/ F12 culture. The differentiated cells were identified by morphological observation and Western blot. Results The cells of the experimental group showed a typical morphology of the neuron-like cells and the control group did not change significantly. Western blot analysis showed that both the experimental group 鈪，

本文編號：2325334