An In Vitro Model for Neuroscience: Differentiation of SH-SY5Y Cells into Cells with Morphological and Biochemical Characteristics of Mature Neurons
An In Vitro Model for Neuroscience: Differentiation of SH-SY5Y Cells into Cells with Morphological and Biochemical Characteristics of Mature Neurons
复制标题
DOI：
10.3233/jad-2010-091363
复制
发表时间：
2010-01-01
影响因子：
4
通讯作者：
Hallbeck, Martin
中科院分区：
文献类型：
--
作者：
Agholme, Lotta;Lindstrom, Tobias;Hallbeck, Martin
神经科学，包括阿尔茨海默病的研究，由于缺乏合适的体外模型来研究人类神经系统而受到阻碍。为了抵消这一点，已经进行了许多将细胞系分化成更多神经元样细胞的尝试，导致神经元特征的部分表达。此外，据报道，神经母细胞瘤细胞系缺乏成熟的tau亚型。我们的目标是开发一种改进的体外模型，产生具有人类成熟神经元的形态和生物化学的可持续细胞。为了获得具有神经元分化和功能的细胞，我们研究了在细胞外基质（ECM）凝胶中结合SH-SY 5 Y细胞的三维培养与据报道具有神经分化作用的几种因素的效果。这导致细胞具有明显的神经元形态，具有长的、广泛分支的神经突。进一步的研究揭示了几种神经特异性标志物的表达，包括突触蛋白Sv 2和核标志物NeuN，以及突触和轴突囊泡运输的存在。此外，这些细胞表达成熟的tau亚型，与未分化的细胞相比，tau蛋白表达显著增加，达到成人大脑中发现的水平。总之，我们发现，用视黄酸预处理，然后进行ECM凝胶培养，结合脑源性神经营养因子、神经调节蛋白β（1）、神经生长因子和维生素D（3）处理，产生了与成年神经元明显相似的可持续细胞。这些细胞还表达具有神经元定位的tau的成人剪接形式，使得这种细胞体外模型在神经科学研究的许多领域，特别是阿尔茨海默病领域中有用。
Neuroscience, including research on Alzheimer's disease, is hampered by the lack of suitable in vitro models to study the human nervous system. To counteract this, many attempts to differentiate cell lines into more neuron-like cells have been performed, resulting in partial expression of neuronal features. Furthermore, it has been reported that neuroblastoma cell lines lack mature isoforms of tau. Our aim was to develop an improved in vitro model, generating sustainable cells with morphology and biochemistry of human, mature neurons. To obtain cells with neuronal differentiation and function, we investigated the effect of combining three-dimensional culturing of SH-SY5Y cells in extracellular matrix (ECM) gel with several factors reported to have neuro-differentiating effects. This resulted in cells with apparent neuronal morphology with long, extensively branched neurites. Further investigation revealed expression of several neurospecific markers including synapse protein Sv2 and nuclear marker NeuN, as well as the presence of synapses and axonal vesicle transport. In addition, these cells expressed mature tau isoforms, and tau protein expression was significantly increased compared to undifferentiated cells, reaching levels found in adult human brain. In conclusion, we found that pre-treatment with retinoic acid followed by ECM gel culturing in combination with brain derived neurotrophic factor, neuregulin beta(1), nerve growth factor, and vitamin D(3) treatment generated sustainable cells with unambiguous resemblance to adult neurons. These cells also expresses adult splicing forms of tau with neuronal localization, making this cellular in vitro model useful in many areas of neuroscience research, particularly the Alzheimer's disease field.