Kristensen B W, Noraberg J, Thiébaud P, Koudelka-Hep M, Zimmer J
Anatomy and Neurobiology, Institute of Medical Biology, SDU-Odense University, Winsløwparken 21, 5000 C, Odense, Denmark.
Brain Res. 2001 Mar 30;896(1-2):1-17. doi: 10.1016/s0006-8993(00)03304-7.
In this study we examined the passive biocompatibility of a three-dimensional microelectrode array (MEA), designed to be coupled to organotypic brain slice cultures for multisite recording of electrophysiological signals. Hippocampal (and corticostriatal) brain slices from 1-week-old (and newborn) rats were grown for 4-8 weeks on the perforated silicon chips with silicon nitride surfaces and 40 microm sized holes and compared with corresponding tissue slices grown on conventional semiporous membranes. In terms of preservation of the basic cellular and connective organization, as visualized by Nissl staining, Timm sulphide silver-staining, microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) immunostaining, the slice cultures grown on chips did not differ from conventionally grown slice cultures. Neither were there any signs of astrogliosis or neurodegeneration around the upper recording part of the 47-microm-high platinum-tip electrodes. Slice cultures grown on a separate set of chips with platinum instead of silicon nitride surfaces also displayed normal MAP2 and GFAP immunostaining. The width of the GFAP-rich zone (glia limitans) at the bottom surface of the slice cultures was the same ( approximately 20 microm) in cultures grown on chips with silicon nitride and platinum surfaces and on conventional insert membranes. The slice cultures grown on chips maintained a normal, subfield differentiated susceptibility to the glutamate receptor agonist N-methyl-D-aspartate (NMDA) and the neurotoxin trimethyltin (TMT), as demonstrated by the cellular uptake of propidium iodide (PI), which was used as a reproducible and quantifiable marker for neuronal degeneration. We conclude that organotypic brain slice cultures can grow on silicon-based three-dimensional microelectrode arrays and develop normally with display of normal subfield differentiated susceptibilities to known excito- and neurotoxins. From this it is anticipated that the set-up, designed for recording of electrophysiological parameters, can be used for long-term studies of defined neuronal networks and provide valuable information on both normal, neurotoxicological and neuropathological conditions.
在本研究中，我们检测了一种三维微电极阵列（MEA）的被动生物相容性，该阵列设计用于与器官型脑片培养物耦合，以进行电生理信号的多部位记录。将1周龄（和新生）大鼠的海马（和皮质纹状体）脑片在具有氮化硅表面和40微米大小孔的穿孔硅芯片上培养4 - 8周，并与在传统半透膜上生长的相应组织切片进行比较。通过尼氏染色、硫代硫化银染色、微管相关蛋白2（MAP2）和胶质纤维酸性蛋白（GFAP）免疫染色观察到，在基本细胞和结缔组织的保存方面，芯片上生长的脑片培养物与传统生长的脑片培养物没有差异。在47微米高的铂尖电极上部记录部分周围，也没有任何星形胶质细胞增生或神经退行性变的迹象。在另一组具有铂而非氮化硅表面的芯片上生长的脑片培养物也显示出正常的MAP2和GFAP免疫染色。在具有氮化硅和铂表面的芯片上以及传统插入膜上生长的培养物中，脑片培养物底面富含GFAP的区域（胶质界膜）宽度相同（约20微米）。如通过碘化丙啶（PI）的细胞摄取所证明的，在芯片上生长的脑片培养物对谷氨酸受体激动剂N - 甲基 - D - 天冬氨酸（NMDA）和神经毒素三甲基锡（TMT）保持正常的、亚区分化的敏感性，碘化丙啶用作神经元变性的可重复和可量化标记。我们得出结论，器官型脑片培养物可以在基于硅的三维微电极阵列上生长并正常发育，对已知的兴奋性毒素和神经毒素表现出正常的亚区分化敏感性。由此预计，为记录电生理参数而设计的装置可用于对特定神经网络的长期研究，并提供有关正常、神经毒理学和神经病理学状况的有价值信息。
