Peng Tao, Zhu Ganghua, Dong Yunpeng, Zeng Junjie, Li Wei, Guo Weiwei, Chen Yong, Duan Maoli, Hocher Berthold, Xie Dinghua
Clin Lab. 2015;61(9):1171-8. doi: 10.7754/clin.lab.2015.150217.
Previous studies have shown that BMP4 may play an important part in the development of auditory neurons (ANs), which are degenerated in sensorineural hearing loss. However, whether BMP4 can promote sensory fate specification from mesenchymal stromal cells (MSCs) is unknown so far.
MSCs isolated from Sprague-Dawley (SD) rats were confirmed by expression of MSC markers using flow cytometry and adipogenesis/osteogenesis using differentiation assays. MSCs treated with a complex of neurotrophic factors (BMP4 group and non-BMP4 group) were induced into auditory neuron-like cells, then the differences between the two groups were analyzed in morphological observation, cell growth curve, qRT-PCR, and immunofluorescence.
Flow cytometric analysis showed that the isolated cells expressed typical MSC surface markers. After adipogenic and osteogenic induction, the cells were stained by oil red O and Alizarin Red. The neuronal induced cells were in the growth plateau and had special forms of neurons. In the presence of BMP4, the inner ear genes NF-M, Neurog1, GluR4, NeuroD, Calretinin, NeuN, Tau, and GATA3 were up-regulated in MSCs.
MSCs have the capacity to differentiate into auditory neuron-like cells in vitro. As an effective inducer, BMP4 may play a key role in transdifferentiation.
先前的研究表明，骨形态发生蛋白4（BMP4）可能在听觉神经元（ANs）的发育中起重要作用，而听觉神经元在感音神经性听力损失中会发生退化。然而，迄今为止，BMP4是否能促进间充质基质细胞（MSCs）向感觉命运的特化尚不清楚。
通过流式细胞术检测MSC标志物的表达以及利用分化试验检测脂肪生成/骨生成，对从Sprague-Dawley（SD）大鼠分离的MSCs进行鉴定。用神经营养因子复合物处理的MSCs（BMP4组和非BMP4组）被诱导分化为听觉神经元样细胞，然后通过形态学观察、细胞生长曲线、qRT-PCR和免疫荧光分析两组之间的差异。
流式细胞术分析表明，分离的细胞表达典型的MSC表面标志物。在进行成脂和成骨诱导后，细胞分别用油红O和茜素红染色。神经元诱导细胞处于生长平台期，具有特殊的神经元形态。在BMP4存在的情况下，内耳基因NF-M、Neurog1、GluR4、NeuroD、钙视网膜蛋白、NeuN、Tau和GATA3在MSCs中上调。
MSCs在体外具有分化为听觉神经元样细胞的能力。作为一种有效的诱导剂，BMP4可能在转分化中起关键作用。
