Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology.
Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology.
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DOI：
10.1007/978-1-62703-640-5_2
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发表时间：
2013
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影响因子：
--
通讯作者：
Langford, Dianne
中科院分区：
文献类型：
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作者：
Kovalevich, Jane;Langford, Dianne
胚胎中枢神经系统组织来源的原代哺乳动物神经元的使用受到以下事实的限制：一旦终末分化为成熟神经元，细胞就不能再繁殖。转化的神经元样细胞系可以在体外使用来克服这一限制。然而，当利用来自恶性肿瘤的细胞时，存在几个警告。在此背景下，流行的SH-SY5Y神经母细胞瘤细胞株及其在体外系统中的应用被描述。SH-SY5Y(ATCC®CRL-2266™)细胞最初来源于转移性骨肿瘤活检，是亲本系SK-N-SH(ATCC®HTB-11™)的子系。将SK-N-SH亚克隆至SH-SY、SH-SY5、SH-SY5Y共3次。SH-SY5Y由比德勒于1970年在ATCC®上存放。在体外研究中使用SH-SY5Y细胞时，应考虑这些细胞的三个重要特征。首先，培养物包括贴壁细胞和漂浮细胞，这两种细胞都是有活力的。很少有研究讨论贴壁和漂浮表型的生物学意义，但大多数报道的研究利用贴壁群体，并在更换培养基时丢弃漂浮细胞。第二，Biedler等人的早期研究表明，亲代分化的SK-N-SH细胞包含两种不同的表型：神经母细胞样细胞和上皮样细胞(Ross等，J Nat Cancer Inst 71：741-747,1983)。这两种表型可能对应于Encinas等人后来在SH-SY5Y研究中描述的“N”和“S”型。(《神经化学杂志》75：991-1003,2000)。具有神经母细胞样形态的细胞呈儿茶酚胺能神经元特有的酪氨酸羟基酶(TH)和多巴胺-β-羟基酶阳性，而上皮样细胞缺乏这些酶活性(Ross等人，《NAT癌症杂志》71：741-747,1983)。第三，SH-SY5Y细胞可以分化为以神经元标志物为特征的更成熟的神经元样表型。有几种方法可以区分SH-SY5Y细胞，如下所述。维甲酸是最常用的分化手段，将详细介绍。
The use of primary mammalian neurons derived from embryonic central nervous system tissue is limited by the fact that once terminally differentiated into mature neurons, the cells can no longer be propagated. Transformed neuronal-like cell lines can be used in vitro to overcome this limitation. However, several caveats exist when utilizing cells derived from malignant tumors. In this context, the popular SH-SY5Y neuroblastoma cell line and its use in in vitro systems is described. Originally derived from a metastatic bone tumor biopsy, SH-SY5Y (ATCC® CRL-2266™) cells are a subline of the parental line SK-N-SH (ATCC® HTB-11™). SK-N-SH were subcloned three times; first to SH-SY, then to SH-SY5, and finally to SH-SY5Y. SH-SY5Y were deposited to the ATCC® in 1970 by June L. Biedler. Three important characteristics of SH-SY5Y cells should be considered when using these cells in in vitro studies. First, cultures include both adherent and floating cells, both types of which are viable. Few studies address the biological significance of the adherent versus floating phenotypes, but most reported studies utilize adherent populations and discard the floating cells during media changes. Second, early studies by Biedler’s group indicated that the parental differentiated SK-N-SH cells contained two morphologically distinct phenotypes: neuroblast-like cells and epithelial-like cells (Ross et al., J Nat Cancer Inst 71:741–747, 1983). These two phenotypes may correspond to the “N” and “S” types described in later studies in SH-SY5Y by Encinas et al. (J Neurochem 75:991–1003, 2000). Cells with neuroblast-like morphology are positive for tyrosine hydroxylase (TH) and dopamine-β-hydroxylase characteristic of catecholaminergic neurons, whereas the epithelial-like counterpart cells lacked these enzymatic activities (Ross et al., J Nat Cancer Inst 71:741–747, 1983). Third, SH-SY5Y cells can be differentiated to a more mature neuron-like phenotype that is characterized by neuronal markers. There are several methods to differentiate SH-SY5Y cells and are mentioned below. Retinoic acid is the most commonly used means for differentiation and will be addressed in detail.