香丹注射液定向诱导大鼠骨髓间质干细胞分化为神经元的特点及其影响因素

Characteristics of Oriented Induction of Xiangdan Injection on Differen tiation o f Marrow Mesenchymal Stem Cells into Neurons and Its Influencing Factors

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中文关键词: 骨髓间质干细胞神经元香丹注射液细胞分化

英文关键词:marrow mesenchymal stem cell neuron Xiangdan injection cell differentiation

基金项目:国家自然科学基金资助项目 (No .30 1 70 337;No.30 370 51 0 ) ;广东省自然科学基金资助项目 (No.970 0 61 ) ;卫生部临床学科重点项目 (No.2 0 0 1 32 1 ) ;博士点专项科研基金 (No .2 0 0 30 5580 58)

作者	单位
黄慧	中山大学附属第一医院神经科
唐云安	四川大学华西公共卫生学院
张成	中山大学附属第一医院神经科

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中文摘要:

目的探讨香丹注射液诱导成年SD大鼠骨髓间质干细胞 (rMSCs)定向分化为神经元样细胞的特点和影响因素。方法体外培养rMSCs至第 5代 ,培养基中加入碱性成纤维生长因子 (bFGF)预诱导2 4h ,更换含不同浓度香丹注射液的诱导液诱导 ,比较香丹注射液rMSCs分化 (诱导率 )在不同的培养基组成、诱导剂浓度和接种时的细胞密度等因素的影响 ,并用台盼蓝染色法、四唑盐比色法 (MTT法 )评判上述因素对诱导后细胞存活率的影响。结果 1%～ 5 %香丹注射液的诱导组 ,均可诱导rMSCs分化为神经元样细胞 ,诱导后 6～ 12h诱导率可达 (83 5± 3 8) % ,90 %以上的细胞存活时间 >36h。其他条件相同时 ,香丹注射液 3%～ 5 %的浓度组、无血清的D/F12 +N2 +bFGF诱导组和 2 5× 10 4 /cm2 的细胞接种密度 ,可得到最高的诱导率和细胞存活率。结论 3%～ 5 %的浓度、无血清D/F12 +N2 +bFGF(10 μg/L)和 2 5× 10 4 /cm2 的细胞接种密度是香丹注射液定向诱导rMSCs分化为神经元样细胞的最适条件。

英文摘要:

ObjectiveTo study the effect of Xiangdan injec tion (XDI) in inducing adult SD rat marrow mesenchymal stem cells (rMSCs) orientedly differentiated i nto neuron-like cells, its characteristics and influencing factors were explore d . MethodsThe 5th generation of rMSCs cultured in vitro were pre-treated for 24 hrs by adding basic fibroblast growth factors (bFGF) into the medium, then th e indu cing liquid was replaced by XDI with different concentration to compare the rMS Cs differentiation rate under different constitution of medium, different concen tration of inducer and cell density of incubation. The induced cell survival rat e under effects of above-mentioned factors was evaluated by trypan blue stain a n d MTT method. ResultsXDI in 1%-5% concentration could induce rMSCs differentia ted into neuron-like cells, the inducing rate reached 83.5±3.8% 6-12 hrs later, more than 90% cells, survival rate was over 36 hrs. The maximal inducing rate a nd cell survival rate could be obtained by treated with 3%-5% XDI, serum-free D/F 12 +N2+bFGF and with the cell density in 2.5×10 4/cm 2, when the other factors were the same. ConclusionXDI of 3%-5% concentration, serum -free D/F12+N2+bFGF (10μg/L) and w ith the cell density incubated of 2.5×10 4/cm 2 is the optimal condition for oriented induction of rMSCs differentiating to neuron-like cells.

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