First human hNT neurons patterned on parylene-C/silicon dioxide substrates: Combining an accessible cell line and robust patterning technology for the study of the pathological adult human brain

Unsworth, Charles P.; Graham, Scott E.; Delivopoulos, Evangelos; Dragunow, Michael; Murray, AF

doi:10.1016/j.jneumeth.2010.09.022

Cited by 33 publications

(34 citation statements)

References 10 publications

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“…The authors have also investigated the protein adhesive properties of parylene-C/SiO 2 substrates. This recently pioneered and patented [13] method has demonstrated that it is possible to accurately pattern primary rat neurons and astrocytes [14] and more recently human neurons [15] on silicon chips, in a reliable and robust manner.…”

Section: Introductionmentioning

confidence: 99%

“…The benchmark parylene-C dimensions reported for successful cultures of primary rat hippocampal neurons and astrocytes are 100 nm thickness and 5e20 mm strip width [14]. Human neurons were also patterned on 30 mm wide and 100 nm thick strips [15]. Capacitive embedded micro-electrode work performed in [17] has demonstrated that extracellular stimulation was possible using a 50 mm diameter spot of a 10 nm insulating layer of oxide for the Retzius cell of the leech (which had the diameter of 70 mm).…”

Section: Introductionmentioning

confidence: 99%

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Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

et al. 2011

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

et al. 2011

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“…Once prepared, activation of chips in fetal bovine serum enables a wide range of cell types to be patterned in culture. Our group has successfully patterned primary murine hippocampal cells [14][15][16] , the HEK 293 cell line 17 , the human neuron-like teratocarcinoma (hNT) cell line 18 , primary murine cerebellar granule cells, and primary human glioma-derived stem-like cells. Figure 4 illustrates the potential to augment the patterning platform by using alternative activation solutions.…”

Section: Protocol Representative Resultsmentioning

confidence: 99%

Cell Patterning on Photolithographically Defined Parylene-C: SiO<sub>2</sub> Substrates

Hughes

Brennan

Bunting

et al. 2014

JoVE

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Cell patterning platforms support broad research goals, such as construction of predefined in vitro neuronal networks and the exploration of certain central aspects of cellular physiology. To easily combine cell patterning with Multi-Electrode Arrays (MEAs) and silicon-based 'lab on a chip' technologies, a microfabrication-compatible protocol is required. We describe a method that utilizes deposition of the polymer parylene-C on SiO 2 wafers. Photolithography enables accurate and reliable patterning of parylene-C at micron-level resolution. Subsequent activation by immersion in fetal bovine serum (or another specific activation solution) results in a substrate in which cultured cells adhere to, or are repulsed by, parylene or SiO 2 regions respectively. This technique has allowed patterning of a broad range of cell types (including primary murine hippocampal cells, HEK 293 cell line, human neuron-like teratocarcinoma cell line, primary murine cerebellar granule cells, and primary human glioma-derived stem-like cells). Interestingly, however, the platform is not universal; reflecting the importance of cell-specific adhesion molecules. This cell patterning process is cost effective, reliable, and importantly can be incorporated into standard microfabrication (chip manufacturing) protocols, paving the way for integration of microelectronic technology.

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“…Although we cannot as yet identify the key protein combinations in serum, we have developed the capacity to induce cell-tolerance to SiO 2 and cell repulsion from parylene (a complete reversal of the serum-induced parylene patterning effects reported previously. [6][7][8][9][10][11] This encourages the use of the parylene/SiO 2 platform for effective cell patterning with a standardized and consistent activation solution and protocol, without recourse to serum (which is poorly characterized and heterogeneous between batches).…”

Section: Rationalized Protein Activation Solutionsmentioning

confidence: 99%

“…10,11 A core motivation for our work is, ultimately, to integrate cell-patterning with either optical or capacitative electrical coupling techniques. This will enable interaction between microelectronic circuits and patterned neuronal networks at a physiologically appropriate spatiotemporal scale.…”

Section: Introductionmentioning

confidence: 99%

Modulating patterned adhesion and repulsion of HEK 293 cells on microengineered parylene‐C/SiO₂ substrates

Hughes

Bunting

Cameron

et al. 2012

J Biomedical Materials Res

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This article describes high resolution patterning of HEK 293 cells on a construct of micropatterned parylene-C and silicon dioxide. Photolithographic patterning of parylene-C on silicon dioxide is an established and consistent process. Activation of patterns by immersion in serum has previously enabled patterning of murine hippocampal neurons and glia, as well as the human hNT cell line. Adapting this protocol we now illustrate high resolution patterning of the HEK 293 cell line. We explore hypotheses that patterning is mediated by transmembrane integrin interactions with differentially absorbed serum proteins, and also by etching the surface substrate with piranha solution. Using rationalized protein activation solutions in place of serum, we show that cell patterning can be modulated or even inverted. These cell-patterning findings assist our wider goal of engineering and interfacing functional neuronal networks via a silicon semiconductor platform.

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First human hNT neurons patterned on parylene-C/silicon dioxide substrates: Combining an accessible cell line and robust patterning technology for the study of the pathological adult human brain

Cited by 33 publications

References 10 publications

Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Cell Patterning on Photolithographically Defined Parylene-C: SiO<sub>2</sub> Substrates

Modulating patterned adhesion and repulsion of HEK 293 cells on microengineered parylene‐C/SiO₂ substrates

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First human hNT neurons patterned on parylene-C/silicon dioxide substrates: Combining an accessible cell line and robust patterning technology for the study of the pathological adult human brain

Cited by 33 publications

References 10 publications

Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Cell Patterning on Photolithographically Defined Parylene-C: SiO<sub>2</sub> Substrates

Modulating patterned adhesion and repulsion of HEK 293 cells on microengineered parylene‐C/SiO2 substrates

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Modulating patterned adhesion and repulsion of HEK 293 cells on microengineered parylene‐C/SiO₂ substrates