Characterization of three human cell line models for high-throughput neuronal cytotoxicity screening.
Characterization of three human cell line models for high-throughput neuronal cytotoxicity screening.
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DOI：
10.1002/jat.3334
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发表时间：
2017-02
影响因子：
3.3
通讯作者：
Gerhold, David
中科院分区：
文献类型：
--
作者：
Tong, Zhi-Bin;Hogberg, Helena;Kuo, David;Sakamuru, Srilatha;Xia, Menghang;Smirnova, Lena;Hartung, Thomas;Gerhold, David
超过7.5万种人造化学物质污染环境；其中许多还没有经过毒性测试。这些化学物质需要定量的高通量筛选分析，以评估它们在神经毒性中的致病作用，包括帕金森氏症和其他神经退行性疾病。为了便于高通量筛选对神经元的细胞毒性，比较了三种人类神经细胞模型：SH-SY5Y神经母细胞瘤细胞、LUHMES条件永生化的多巴胺能神经元和来自人胎脑的神经干细胞(NSC)。评估了这三种细胞系的分化速度和程度，以及对32种已知或候选神经毒物的敏感性。首先，在7天的分化期内检测神经分化基因的表达。在三种细胞系中，LUHMES分化后神经元标志物的基因表达水平最高。在未分化状态和分化7天后，LUHMES细胞对大多数疑似或已知的32种神经毒物表现出比SH-SY5Y或NSCs更高的细胞毒敏感性。LUHMES细胞的独特之处还在于，与未分化状态相比，分化状态下的LUHMES细胞对几种化合物更敏感；包括已知的神经毒物秋水仙碱、甲基汞(II)和长春新碱。基因表达结果表明，分化中的LUHMES细胞可能因为表达低水平的抗凋亡基因BCL2和BIRC5/Survivin而对凋亡敏感，而SH-SY5Y细胞可能因为高水平表达BCL2、BIRC5/Survivin和BIRC3基因而抵抗凋亡。因此，LUHMES细胞表现出良好的神经细胞毒性筛选特性：快速分化为高表达神经元标记基因的神经元，以及LUHMES细胞对已知神经毒物的显著敏感性。三种人类神经细胞系被评估为神经元细胞毒性的高通量筛选模型：SH-SY5Y神经母细胞瘤细胞、LUHMES条件永生化的多巴胺能神经元和神经干细胞。分化7天后，LUHMES表达的神经元标志物水平最高。与已分化的SH-SY5Y或NSCs相比，分化的LUHMES细胞对大多数疑似或已知的32种神经毒物表现出更高的细胞毒敏感性，对11种化合物的细胞毒敏感性比未分化的LUHMES细胞更高。
More than 75,000 man-made chemicals contaminate the environment; many of these have not been tested for toxicities. These chemicals demand quantitative high-throughput screening assays to assess them for causative roles in neurotoxicities, including Parkinson’s disease and other neurodegenerative disorders. To facilitate high throughput screening for cytotoxicity to neurons, three human neuronal cellular models were compared: SH-SY5Y neuroblastoma cells, LUHMES conditionally-immortalized dopaminergic neurons, and Neural Stem Cells (NSC) from human fetal brain. These three cell lines were evaluated for rapidity and degree of differentiation, and sensitivity to 32 known or candidate neurotoxicants. First, expression of neural differentiation genes was assayed during a 7-day differentiation period. Of the three cell lines, LUHMES showed the highest gene expression of neuronal markers after differentiation. Both in the undifferentiated state and after 7 days of neuronal differentiation, LUHMES cells exhibited greater cytotoxic sensitivity to most of 32 suspected or known neurotoxicants than SH-SY5Y or NSCs. LUHMES cells were also unique in being more susceptible to several compounds in the differentiating state than the undifferentiated state; including known neurotoxicants colchicine, methyl-mercury (II), and vincristine. Gene expression results suggest that differentiating LUHMES cells may be susceptible to apoptosis because they express low levels of anti-apoptotic genes BCL2 and BIRC5/survivin; whereas SH-SY5Y cells may be resistant to apoptosis because they express high levels of BCL2, BIRC5/survivin, and BIRC3 genes. Thus, LUHMES cells exhibited favorable characteristics for neuro-cytotoxicity screening: rapid differentiation into neurons exhibiting high level expression neuronal marker genes, and marked sensitivity of LUHMES cells to known neurotoxicants. Three human neuronal cell lines were evaluated as high throughput screening models for neuronal cytotoxicity: SH-SY5Y neuroblastoma cells, LUHMES conditionally-immortalized dopaminergic neurons, and Neural Stem Cells. After 7 days of differentiation LUHMES expressed the highest levels of neuronal markers. Differentiated LUHMES cells exhibited greater cytotoxic sensitivity to most of 32 suspected or known neurotoxicants than differentiated SH-SY5Y or NSCs, and greater cytotoxic sensitivity to 11 compounds compared to undifferentiated LUHMES cells.
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影响因子：
3.4
作者：
Cheung, Yuen-Ting;Lau, Way Kwok-Wai;Chang, Raymond Chuen-Chung
通讯作者：
Chang, Raymond Chuen-Chung
影响因子：
4.1
作者：
Huang, Ruili;Southall, Noel;Austin, Christopher P.
通讯作者：
Austin, Christopher P.
影响因子：
2.6
作者：
Hsu, Yi-Chiang;Chang, Sue-Joan;Huang, Tzuu-Yuan
通讯作者：
Huang, Tzuu-Yuan
DOI：
10.1023/a:1027324230923
发表时间：
2003-01-01
期刊：
JOURNAL OF NEUROCYTOLOGY
影响因子：
--
作者：
Buntinx, M;Vanderlocht, J;Steels, P
通讯作者：
Steels, P
影响因子：
4.5
作者：
IKEGWUONU, FI
通讯作者：
IKEGWUONU, FI