体外诱导大鼠骨髓间充质干细胞向GABA能神经元分化
发布时间：2018-11-11 15:47
【摘要】：目的研究骨髓间充质干细胞(BMSCs)在体外向γ-氨基丁酸(GABA)能神经元方向的分化。方法大鼠股骨骨髓分离出BMSCs,培养传代,通过10%胎牛血清(FBS)、20 ng/ml表皮生长因子(EGF)和改良Eagle培养基(DMEM/F12)预诱导24 h,随之加入含10 ng/ml碱性成纤维细胞生长因子(bFGF)、10 ng/ml骨形成蛋白-2(BMP-2)和10 ng/ml脑源性神经营养因子(BDNF)作用48 h后,再以10μmol/L全反式维甲酸(ATRA)培养48 h,最后用40 mmol/L KCl刺激15 min完成诱导分化。对照组用10%FBS和DMEM/F12培养不添加任何诱导因子。形态学观察和Western blot对分化后的细胞进行鉴定分析。结果实验组细胞在形态学上表现出典型的神经元样细胞形态,对照组无明显变化;Western blot分析知实验组Ⅰ和实验组Ⅱ都有分化为GABA能神经元的趋势,但实验组Ⅱ的分化率高于实验组Ⅰ。结论BMSCs可在体外分化为GABA能神经元。
[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，
本文编号：2325334
[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，
本文编号：2325334
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2325334.html
最近更新
教材专著
