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神经元细胞生长和分化对硅纳米线晶体管界面效应的非法拉第电阻抗研究。

Non-Faradaic electrical impedimetric investigation of the interfacial effects of neuronal cell growth and differentiation on silicon nanowire transistors.

作者信息

Lin Shu-Ping, Vinzons Lester U, Kang Yu-Shan, Lai Tung-Yen

机构信息

†Graduate Institute of Biomedical Engineering, National Chung Hsing University, Taichung 40227, Taiwan, R.O.C.

‡National Nano Device Laboratories, Hsinchu 30078, Taiwan, R.O.C.

出版信息

ACS Appl Mater Interfaces. 2015 May 13;7(18):9866-78. doi: 10.1021/acsami.5b01878. Epub 2015 May 1.

Abstract

Silicon nanowire field-effect transistor (SiNW FET) devices have been interfaced with cells; however, their application for noninvasive, real-time monitoring of interfacial effects during cell growth and differentiation on SiNW has not been fully explored. Here, we cultured rat adrenal pheochromocytoma (PC12) cells, a type of neural progenitor cell, directly on SiNW FET devices to monitor cell adhesion during growth and morphological changes during neuronal differentiation for a period of 5-7 d. Monitoring was performed by measuring the non-Faradaic electrical impedance of the cell-SiNW FET system using a precision LCR meter. Our SiNW FET devices exhibited changes in impedance parameters during cell growth and differentiation because of the negatively charged cell membrane, seal resistance, and membrane capacitance at the cell/SiNW interface. It was observed that during both PC12 cell growth and neuronal differentiation, the impedance magnitude increased and the phase shifted to more negative values. However, impedance changes during cell growth already plateaued 3 d after seeding, while impedance changes continued until the last observation day during differentiation. Our results also indicate that the frequency shift to above 40 kHz after growth factor induction resulted from a larger coverage of cell membrane on the SiNWs due to distinctive morphological changes according to vinculin staining. Encapsulation of PC12 cells in a hydrogel scaffold resulted in a lack of trend in impedance parameters and confirmed that impedance changes were due to the cells. Moreover, cytolysis of the differentiated PC12 cells led to significant changes in impedance parameters. Equivalent electrical circuits were used to analyze the changes in impedance values during cell growth and differentiation. The technique employed in this study can provide a platform for performing investigations of growth-factor-induced progenitor cell differentiation.

摘要

硅纳米线场效应晶体管(SiNW FET)器件已与细胞连接;然而,其在非侵入性实时监测细胞在硅纳米线上生长和分化过程中的界面效应方面的应用尚未得到充分探索。在这里,我们将大鼠肾上腺嗜铬细胞瘤(PC12)细胞(一种神经祖细胞)直接培养在SiNW FET器件上,以监测其生长过程中的细胞黏附以及神经元分化过程中的形态变化,为期5 - 7天。通过使用精密LCR仪表测量细胞 - SiNW FET系统的非法拉第电阻抗来进行监测。由于细胞/硅纳米线界面处带负电的细胞膜、密封电阻和膜电容,我们的SiNW FET器件在细胞生长和分化过程中呈现出阻抗参数的变化。观察到在PC12细胞生长和神经元分化过程中,阻抗幅值均增加且相位移向更负值。然而,细胞生长过程中的阻抗变化在接种后3天就已趋于平稳,而在分化过程中阻抗变化一直持续到最后观察日。我们的结果还表明,生长因子诱导后频率移至40 kHz以上是由于根据纽蛋白染色观察到的独特形态变化,导致细胞膜在硅纳米线上的覆盖面积更大。将PC12细胞封装在水凝胶支架中导致阻抗参数缺乏变化趋势,并证实阻抗变化是由细胞引起的。此外,分化后的PC12细胞的细胞溶解导致阻抗参数发生显著变化。使用等效电路来分析细胞生长和分化过程中阻抗值的变化。本研究中采用的技术可为开展生长因子诱导的祖细胞分化研究提供一个平台。

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