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SH-SY5Y

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SH-SY5Y细胞可以形成未分化的细胞簇,然后将分化细胞传播到周围区域。 这种情况被称为“过度容易形成”(over-easy formation)。

SH-SY5Y是一种常应用于科学研究的人类神经母细胞瘤细胞系,最初是从一名拥有成神经母细胞瘤的人类四岁女童的骨髓活检组织中分离出来的,具有两个X染色体且没有Y染色体,因为该细胞系来自于一个人类女童。SH-SY5Y细胞具有肾上腺素能表型(adrenergic phenotype),但同时也表达着多巴胺能标记(dopaminergic markers)。目前已知SH-SY5Y细胞具有中等多巴胺β羟化酶活性[1],而胆碱乙酰转移酶英语Choline acetyltransferase乙酰胆碱酯酶丁酰胆碱酯酶英语Butyrylcholinesterase水平则可以忽略不计,并且发现SH-SY5Y细胞具有基础水平的去甲肾上腺素释放[2]和酪氨酸羟化酶活性[3]

SH-SY5Y细胞通常用作神经元功能和细胞分化体外模型。目前已被用于研究神经科学的多个领域,包括帕金森氏症[4]阿兹海默症神经毒性、局部缺血肌萎缩性脊髓侧索硬化症的研究等[5][6][7] ,甚至可以用于研究脑细胞的其他特征和神经发生[8]

除此之外,SH-SY5Y细胞也常用于研究α-突触核蛋白英语Alpha-synuclein,因为未转染的SH-SY5Y细胞中有自发的α-突触核蛋白聚集现象[9],而且发现已分化和未分化的SH-SY5Y细胞均对细胞外α-突触核蛋白诱导的毒性敏感[10],因此已用于研究α-突触核蛋白降解的动力学和机理[11][12]、α-突触核蛋白聚集与细胞内之间的联系[13],以及α-突触核蛋白的其他病理变化,例如其羧基末端的裂解[14]

历史

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SH-SY5Y细胞是从骨髓活检衍生品系SK-N-SH细胞中克隆出来的[15],而克隆过程涉及选择表达神经元样特征的单个细胞或细胞簇。 SK-N-SH细胞的成神经细胞样亚克隆被命名为SH-SY细胞,后来被亚克隆为SH-SY5细胞,接着再进行第三次亚克隆以产生SH-SY5Y细胞,最早于1978年报导[1]

形态学

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SH-SY5Y细胞通常以两种不同的方式在组织培养中生长。第一种方式是长成团块的细胞漂浮在培养基中,而第二种方式是长成团块的细胞粘在碟子上。SH-SY5Y细胞可以在体外自发地在两种表型之间转化,即成神经细胞样细胞和上皮样细胞,尽管目前尚不清楚该过程的机制。然而,鉴于其形态及拥有将细胞分化为神经元系的能力,该细胞系被认为是N型。这与同样来自SK-N-SH细胞的S型亚克隆细胞系SH-EP细胞相反[16] 。将具有类神经突的短刺状细胞从这些粘附的团块中移出,并且进行观察后会发现SH-SY5Y细胞具有异常的1号染色体,当中有一个1q节的副本,而该副本被称为1q染色体三倍体(trisomy 1q)。SH-SY5Y细胞既通过有丝分裂繁殖,又通过将神经突延伸到周围区域而分化。细胞分裂时,聚集的细胞看上去与分化后的细胞有很大的不同,以至于刚入行的科学家经常将此现象视为细胞被污染。分裂中的SH-SY5Y细胞可以形成细胞簇,提醒它们的癌性,但是某些使用到视黄酸脑源性神经营养因子tPA的治疗方法会令细胞树突状化及分化。此外,视黄酸诱导可以抑制细胞生长,并且增加SH-SY5Y细胞中去甲肾上腺素的产生[17][18]

细胞培养

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目前最常使用的培养方式是伊格尔最低限度必需培养基(DMEM)和Ham's F-12培养基英语Ham's tissue culture medium的1:1比例混合物,以及10%补充性胎牛血清。DMEM通常包含1.5g/L 碳酸氢钠、2.0 mM L-麸酰胺酸、1.0 mM丙酮酸钠,以及0.1 mM非必需氨基酸。SH-SY5Y细胞必要在37℃、95%的空气和5%的二氧化碳的环境下生长,并且被建议在烧瓶中培养SH-SY5Y细胞,而烧瓶已涂上有利于细胞粘附的涂层。这将有助于神经母细胞瘤的细胞分化和树突化。这些细胞通常非常顽强,可以在最广泛使用的培养基中生长。随着细胞传代次数的增加,神经元特征会出现丧失,故而被建议不要在第20代后,使用或验证去甲肾上腺素的摄取,或者神经元肿瘤标志物等特定特征。除此之外,为了获得具有神经元表型的衍生细胞,已经有许多报道描述了多种的分化方案[19][20][21][22][23]。早期对SH-SY5Y细胞的研究表明,利用星形孢菌素可使其分化为神经元表型[24][25],后来被表征为儿茶酚胺[26]。因此,应该仔细表征不同浓度的星形孢菌素对SH-SY5Y细胞的作用,以正确解释将星形孢菌素用作分化剂或凋亡诱导剂的研究[4]

参考资料

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外部链接

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