Liu Bin, Deng Chunying, Zhang Yuqin, Zhang Jinxia
First Department of Neurology, Hospital Affiliated to Hebei United University, Tangshan 063000, Hebei Province, China.
Neural Regen Res. 2012 Jul 5;7(19):1463-8. doi: 10.3969/j.issn.1673-5374.2012.19.003.
The present study analyzed changes in Wnt3a expression during differentiation of adipose-derived stem cells into cholinergic neurons. Immunocytochemistry and immunofluorescence revealed significantly increased nestin, neuron-specific enolase, microtubule-associated protein 2, and choline acetyltransferase expression in adipose-derived stem cells isolated from Sprague-Dawley rats and cultured in vitro in neural-induced medium. These expressions increased with prolonged induction time. Real-time reverse transcription-PCR and western blot assay results demonstrated significantly increased choline acetyltransferase and Wnt3a protein and mRNA expressions, respectively, in adipose-derived stem cells following induction. Choline acetyltransferase expression positively correlated with Wnt3a protein and mRNA expressions. These results demonstrated that neural-induced medium induced differentiation of adipose-derived stem cells into cholinergic neuronal-like cells, with subsequent increased Wnt3a expression.
本研究分析了脂肪干细胞向胆碱能神经元分化过程中Wnt3a表达的变化。免疫细胞化学和免疫荧光显示,从Sprague-Dawley大鼠分离并在神经诱导培养基中体外培养的脂肪干细胞中,巢蛋白、神经元特异性烯醇化酶、微管相关蛋白2和胆碱乙酰转移酶的表达显著增加。这些表达随着诱导时间的延长而增加。实时逆转录-PCR和蛋白质印迹分析结果表明,诱导后脂肪干细胞中胆碱乙酰转移酶和Wnt3a蛋白及mRNA表达分别显著增加。胆碱乙酰转移酶表达与Wnt3a蛋白及mRNA表达呈正相关。这些结果表明,神经诱导培养基可诱导脂肪干细胞分化为胆碱能神经元样细胞,并随后增加Wnt3a表达。