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利用新型阻抗生物传感器研究神经元分化和信号转导过程。

Studies on neuronal differentiation and signalling processes with a novel impedimetric biosensor.

机构信息

Faculty of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece.

出版信息

Biosens Bioelectron. 2010 Dec 15;26(4):1407-13. doi: 10.1016/j.bios.2010.07.066. Epub 2010 Jul 23.

Abstract

The differentiation of neural cells is an important process during the development of the central nervous system. Studies on the mechanisms of the differentiation process is of special importance, e.g. in the field of regenerative medicine. In this contribution the cellular differentiation of gel matrix embedded neuronal cells was studied. The three-dimensional organization of neuronal cells represents a new cellular model system closer to the physiology than conventional two-dimensional cell cultures. Neuro2a (N2a) neuroblastoma cells were immobilized in different gel matrices and the grade of differentiation was compared. Furthermore, the impact of the cell number and selected differentiation factors were analyzed. Experimental results revealed that gel matrices based on collagen-laminin mixtures in contact with serum free medium enable neural differentiation. Therefore, collagen-laminin gels appear as a suitable three-dimensional model for drug screening in developmental neurobiology. Following optimization of the immobilization process, a novel impedimetric sensor and electrical impedance spectroscopy technique was applied to on-line monitor the differentiation process by means of changes in the dielectric and conductive properties. Experimental results showed an increase in the impedance magnitude that can be mainly attributed to differentiating cells accompanied by an increase in the specific resistivity of the bare gel mixture.

摘要

神经细胞的分化是中枢神经系统发育过程中的一个重要过程。对分化过程机制的研究具有特殊意义,例如在再生医学领域。本研究探讨了凝胶基质包埋神经元细胞的细胞分化。神经元细胞的三维组织代表了一种新的细胞模型系统,比传统的二维细胞培养更接近生理学。将神经母细胞瘤细胞(Neuro2a,N2a)固定在不同的凝胶基质中,并比较其分化程度。此外,还分析了细胞数量和所选分化因子的影响。实验结果表明,与无血清培养基接触的基于胶原-层粘连蛋白混合物的凝胶基质能够促进神经分化。因此,胶原-层粘连蛋白凝胶作为发育神经生物学中药物筛选的合适三维模型。在优化固定化过程后,采用新型阻抗传感器和电阻抗谱技术,通过介电和导电特性的变化来在线监测分化过程。实验结果表明,阻抗幅度增加,这主要归因于伴随裸凝胶混合物比电阻增加的分化细胞。

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