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为确定诱导大鼠骨髓基质细胞(BMSCs)向神经元样细胞转分化的适宜培养代数,本研究比较了不同培养代数的BMSCs的转分化潜能。首先体外培养大鼠BMSCs并传代,于不同的培养代数利用碱性成纤维细胞生长因子(bFGF)、神经生长因子(NGF)、全反式维甲酸(RA)、音猬因子(Shh)诱导分化,随后运用免疫荧光法和免疫印迹法检测诱导细胞中的神经元样细胞。结果显示骨髓基质细胞体外培养1至3代时能够被诱导分化为神经元样细胞,培养至第7代时丧失转分化为神经元样细胞的潜能。上述结果提示:传代次数增多将引起BMSCs自发分化,使其向神经元样细胞转分化的能力下降,因而BMSCs培养至第3代时适宜进行诱导分化。
In order to determine suitable culture algebra to induce BMSCs to transdifferentiate into neuron-like cells, we compared the transdifferentiation potential of BMSCs with different culture algebra. First, rat BMSCs were cultured and passaged in vitro. Differentiation was induced by basic fibroblast growth factor (bFGF), nerve growth factor (NGF), all-trans retinoic acid (RA), and sonic hedgehog (Shh) , Followed by immunofluorescence and immunoblotting to detect neuronal-like cells in the induced cells. The results showed that bone marrow stromal cells can be induced to differentiate into neuron-like cells when they were cultured for 1-3 generations in vitro and lose the potential of transdifferentiation into neuron-like cells up to the seventh generation. The above results suggest that: the increased number of passages will cause spontaneous differentiation of BMSCs, so that the ability to transdifferentiate into neuron-like cells decreased, so the BMSCs cultured to the third generation suitable for differentiation.