Studying the formation of large cell aggregates in patterned neuronal cultures

Zeng, Huan-Chang; Ho, Yi Cheng; Chen, Shih-Ta; Wu, Huei-In; Tung, Hsi-Wen; Fang, Weileun; Chang, Y. C.

doi:10.1016/j.jneumeth.2007.05.027

Cited by 4 publications

(3 citation statements)

References 30 publications

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“…Established techniques in the field include micro-contact stamping [1] and microfluidics [2]. Thin film micro-contact printing [3], and use of PDMS or parylene-C stencils [4], [5] are recently developed methods. Patterned neuronal cultures [6] are constantly employed in multi electrode arrays (MEA) [7], [8] and electrophysiological recordings, not only to understand the in vivo development of such networks, but also to investigate how information is processed in the brain [9].…”

Section: Introductionmentioning

confidence: 99%

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Delivopoulos

Murray²

2011

PLoS ONE

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This paper explores the long term development of networks of glia and neurons on patterns of Parylene-C on a SiO2 substrate. We harvested glia and neurons from the Sprague-Dawley (P1–P7) rat hippocampus and utilized an established cell patterning technique in order to investigate cellular migration, over the course of 3 weeks. This work demonstrates that uncontrolled glial mitosis gradually disrupts cellular patterns that are established early during culture. This effect is not attributed to a loss of protein from the Parylene-C surface, as nitrogen levels on the substrate remain stable over 3 weeks. The inclusion of the anti-mitotic cytarabine (Ara-C) in the culture medium moderates glial division and thus, adequately preserves initial glial and neuronal conformity to underlying patterns. Neuronal apoptosis, often associated with the use of Ara-C, is mitigated by the addition of brain derived neurotrophic factor (BDNF). We believe that with the right combination of glial inhibitors and neuronal promoters, the Parylene-C based cell patterning method can generate structured, active neural networks that can be sustained and investigated over extended periods of time. To our knowledge this is the first report on the concurrent application of Ara-C and BDNF on patterned cell cultures.

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Section: Introductionmentioning

confidence: 99%

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Delivopoulos

Murray²

2011

PLoS ONE

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“…This system can then be used as a tool to investigate and identify the signaling molecules involved in cell behaviour and function. For example, surface patterns of cell adhesive peptides or concentration gradients of signaling molecules have influenced cell elongation, formation of neuron aggregates, and controlled axon guidance in 2D cultures Henley and Poo, 2004;Karp et al, 2006;Lohof et al, 1992;Yuan et al, 2003;Zeng et al, 2007). Findings from these studies can enhance our understanding of developmental diseases and design strategies in regenerative medicine.…”

Section: Introductionmentioning

confidence: 99%

Miniaturized system of neurotrophin patterning for guided regeneration

Wosnick

Shoichet

2008

Journal of Neuroscience Methods

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“…As can be observed from Figure 3D,E, after 14 DIV, the cells formed an intricated network spreading over the entire active area surface. In particular, the cell culture protocol (specifically the coating and seeding density) was optimized to limit the clustering phenomena that can be observed in aged neuronal cultures and which may interfere with the culture spatial arrangement (Zeng et al, 2007). We also preformed SEM on cultured neurons on chip (3DIV), following the procedure described in Section “Scanning Electron Microscopy Characterization.” Figure 3F shows extended neuronal networks over an individual active area and with larger magnification on a smaller region (inset).…”

Section: Resultsmentioning

confidence: 99%

High-Density Electrical Recording and Impedance Imaging With a Multi-Modal CMOS Multi-Electrode Array Chip

et al. 2019

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Multi-electrode arrays, both active or passive, emerged as ideal technologies to unveil intricated electrophysiological dynamics of cells and tissues. Active MEAs, designed using complementary metal oxide semiconductor technology (CMOS), stand over passive devices thanks to the possibility of achieving single-cell resolution, the reduced electrode size, the reduced crosstalk and the higher functionality and portability. Nevertheless, most of the reported CMOS MEA systems mainly rely on a single operational modality, which strongly hampers the applicability range of a single device. This can be a limiting factor considering that most biological and electrophysiological dynamics are often based on the synergy of multiple and complex mechanisms acting from different angles on the same phenomena. Here, we designed a CMOS MEA chip with 16,384 titanium nitride electrodes, 6 independent operational modalities and 1,024 parallel recording channels for neuro-electrophysiological studies. Sixteen independent active areas are patterned on the chip surface forming a 4 × 4 matrix, each one including 1,024 electrodes. Electrodes of four different sizes are present on the chip surface, ranging from 2.5 × 3.5 μm 2 up to 11 × 11.0 μm 2 , with 15 μm pitch. In this paper, we exploited the impedance monitoring and voltage recording modalities not only to monitor the growth and development of primary rat hippocampal neurons, but also to assess their electrophysiological activity over time showing a mean spike amplitude of 144.8 ± 84.6 μV. Fixed frequency (1 kHz) and high sampling rate (30 kHz) impedance measurements were used to evaluate the cellular adhesion and growth on the chip surface. Thanks to the high-density configuration of the electrodes, as well as their dimension and pitch, the chip can appreciate the evolutions of the cell culture morphology starting from the moment of the seeding up to mature culture conditions. The measurements were confirmed by fluorescent staining. The effect of the different electrode sizes on the spike amplitudes and noise were also discussed. The multi-modality of the presented CMOS MEA allows for the simultaneous assessment of different physiological properties of the cultured neurons. Therefore, it can pave the way both to answer complex fundamental neuroscience questions as well as to aid the current drug-development paradigm.

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Studying the formation of large cell aggregates in patterned neuronal cultures

Cited by 4 publications

References 30 publications

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Miniaturized system of neurotrophin patterning for guided regeneration

High-Density Electrical Recording and Impedance Imaging With a Multi-Modal CMOS Multi-Electrode Array Chip

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