Nanoscaffold's stiffness affects primary cortical cell network formation

Abstract

Networks of neurons cultured on-chip can provide insights into both normal and disease-state brain function. The ability to guide neuronal growth in specific, artificially designed patterns allows us to study how brain function follows form. Primary cortical cells cultured on nanograting scaffolds, in particular astrocytes, showed highly ordered regions of dendritic outgrowth. Usually, materials suitable for nanopatterning have a stiffness far above that of the extracellular matrix. In this paper, the authors studied two materials with large differences in stiffness, polydimethylsiloxane (PDMS) and silicon. Our results show that both nanopatterned silicon and PDMS guide the outgrowth of astrocytes in cortical cell culture, but the growth of the astrocyte is affected by the stiffness of the substrate, as revealed by differences in the cell soma size and the organization of the outgrowth.

Nanoscale membrane actuator for in vitro mechano-stimuli responsive studies of neuronal cell networks on chip

“…It has been demonstrated that the linear nanogrooved pattern ('nanoscaffold' [35]) on the PDMS surface influences the direction of the neuronal outgrowth as well as the organisation of the cortical neuronal cell networks formation in our previous work [35,44]. Contrastive to the random neuronal network of the primary cortical cells on a non-patterned surface, the neuronal cell network formed on a linear patterned surface is organised with highly ordered cell outgrowths which are aligned with the direction of the line pattern of the nanogrooves.…”

“…Contrastive to the random neuronal network of the primary cortical cells on a non-patterned surface, the neuronal cell network formed on a linear patterned surface is organised with highly ordered cell outgrowths which are aligned with the direction of the line pattern of the nanogrooves. The latter has been revealed by both live/ dead staining and fluorescent immunostaining of the cortical cells [35,44]. Here, Ca 2+ was used to further visualise the dynamic activities in the live neuronal cell network.…”

Advances in 3D neuronal cell culture

“…Materials and the fabrication process details are described in our previous work. 30,31 In brief, the patterning process contains the following steps: the scaffold was first fabricated by dispensing and imprinting the resist using the Imprio55 equipment (Molecular Imprints, Inc., USA) with a quartz stamp containing nanogrooved features on a silicon wafer precoated with bottom antireflective coating layer. Subsequently, the resist scaffold was copied by spin coating a PDMS layer with 100 lm thickness and curing the PDMS at 80 C for 30 min.…”

“…30 and 31) whereas matrix stiffness can impact the phenotype and genotype of neuronal cells. 31,32 In addition, mechanically driven cell polarization in brain tissues and neurotherapeutic approaches using functionalized supermagnetic nanoparticles to potentially restore disordered neural circuits have also been investigated. 33 In order to achieve such a brain on a chip construct ( Fig.…”

Displacement Talbot lithography nanopatterned microsieve array for directional neuronal network formation in brain-on-chip

“…We have also previously demonstrated that a surface with nanopatterns, in particular, nanogrooves, can provide topographical cues to guide cell outgrowth in a primary cortical cell network influencing its organization. 13,14 In this paper, we combined the 3D microsieving structure with nanogrooves to develop a nanopatterned microsieve array for achieving a highly guided cell network in culture. Here, displacement Talbot lithography (DTL) was utilized to pattern the planar surface between the pores of the microsieve because of its ability to create submicrometer sized features at high speed (compared to other nanolithography techniques) and thus low cost, as well as its advantage of providing good patterning uniformity atop of nonflat surfaces.…”