Human Co-culture Model of Neurons and Astrocytes to Test Acute Cytotoxicity of Neurotoxic Compounds
Human Co-culture Model of Neurons and Astrocytes to Test Acute Cytotoxicity of Neurotoxic Compounds
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DOI：
10.1177/1091581817739428
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发表时间：
2017-11-01
影响因子：
2.2
通讯作者：
Coccini, Teresa
中科院分区：
文献类型：
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作者：
De Simone, Uliana;Caloni, Francesca;Coccini, Teresa
对现代毒理学家来说，替代方法及其在计划和开展毒理学实验中的应用已经变得至关重要，从而减少或取代了活的动物。尽管体外人类共培养模型允许建立生物相关的细胞-细胞相互作用，概括组织微环境并更好地模拟其生理，但专门针对这一科学领域和利用这种测试方法可以为毒理学研究提供额外有价值的模型的出版物数量有限。本研究采用人神经细胞（SH-SY5Y细胞系）与胶质细胞即星形胶质细胞（D384细胞系）结合的transwell系统，建立了基于中枢神经系统（CNS）细胞共培养的体外模型，研究D384对SH-SY5Y的神经保护作用，反之亦然。采用该模型检测3种不同神经毒物的急性（24-48小时）细胞毒性：(1)甲基汞（1-2.5 M）， (2) Fe3O4纳米颗粒（1-100 g/mL），(3)甲基乙二醛（0.5-1 mM）。将数据与单培养进行比较，评估线粒体功能和细胞形态。结果清楚地表明，在单培养和共培养条件下，所有测试的化合物都影响线粒体活性和细胞形态。然而，当星形胶质细胞与神经元一起培养时，会减弱单独培养的神经元所产生的神经毒物诱导的细胞毒性作用，并且星形胶质细胞在神经元存在时变得更具抵抗力。这种人类中枢神经系统共培养系统似乎是一种适合的细胞模型，用于高通量急性筛选平台，以及评估人类神经元和星形胶质细胞的毒性和新兴材料（如纳米材料）的神经保护作用，以及由于功能性神经元-星形胶质细胞代谢相互作用而提高灵敏度的新产品。
Alternative methods and their use in planning and conducting toxicology experiments have become essential for modern toxicologists, thus reducing or replacing living animals. Although in vitro human co-culture models allow the establishment of biologically relevant cell-cell interactions that recapitulate the tissue microenvironment and better mimic its physiology, the number of publications is limited specifically addressing this scientific area and utilizing this test method which could provide an additional valuable model in toxicological studies. In the present study, an in vitro model based on central nervous system (CNS) cell co-cultures was implemented using a transwell system combining human neuronal cells (SH-SY5Y cell line) and glial cells, namely astrocytes (D384 cell line), to investigate neuroprotection of D384 on SH-SY5Y and vice versa. The model was applied to test acute (24-48 hours) cytotoxicity of 3 different neurotoxicants: (1) methyl mercury (1-2.5 M), (2) Fe3O4 nanoparticles (1-100 g/mL), and (3) methylglyoxal (0.5-1 mM). Data were compared to mono-cultures evaluating the mitochondrial function and cell morphology. The results clearly showed that all compounds tested affected the mitochondrial activity and cell morphology in both mono-culture and co-culture conditions. However, astrocytes, when cultured together with neurons, diminish the neurotoxicant-induced cytotoxic effects that occurred in neurons cultured alone, and astrocytes become more resistant in the presence of neurons. This human CNS co-culture system seems a suitable cell model to feed high-throughput acute screening platforms and to evaluate both human neuronal and astrocytic toxicity and neuroprotective effects of new and emerging materials (eg, nanomaterials) and new products with improved sensitivity due to the functional neuron-astrocyte metabolic interactions.