Cell Patterning on Photolithographically Defined Parylene-C: SiO₂ Substrates

Abstract

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.

Light-Induced Cell Alignment and Harvest for Anisotropic Cell Sheet Technology

“…Though cell patterning on TiO 2 film have been reported previously, − we here described another novel method to generate micropatterned cues directly on TiO 2 surface without the cell-repellent coating. We observed the effects of UV pretreatment at aspects of wettability, protein adsorption and cell attachment to confirm the generation of micropatterned cues, as shown in Figure .…”

“…Recently, light patterning has been demonstrated as a way to fabricate the hydrophilic/hydrophobic patterns, micropatterned polymer brushes, and small-molecule organic nanowire on TiO 2 film. A novel method for fabricating cell microarrays based on photocatalysis of cell-repellent coating on TiO 2 film was also reported. − In fact, one property of TiO 2 , known as photofunctionalization, showed the great improvements in the attachment and proliferation of osteogenic cells on ultraviolet (UV) light preirradiated TiO 2 film. − Surface hydroxyl group features on TiO 2 controlled by photofunctionalization initiate a hierarchical signal processing thus guide the selective protein adsorption as well as the subsequent cell behaviors. ,, It implied the possibility of cell patterning by UV light on TiO 2 film without the cell-repellent coating and the combination with light-induced cell detachment. Meanwhile, the newly developed photopolymerizable gelatin methacrylate (GelMA), synthesized by adding methacrylate groups to amine-containing side groups of gelatin, , might be an ideal photo-cross-linkable hydrogel to integrate detachment and stack of the light-induced CS.…”

Cost-efficient boundary-free surface patterning achieves high effective-throughput of time-lapse microscopy experiments

“…But such effort is not compatible with all types of cell experiment, as the physical confinement and contact induce phenotypic changes [19][20][21]. Furthermore, photolithography is another approach to deposit either physical or non-physical boundaries for cell confinement [22][23][24]. However, the high cost of this approach distances it from being widely used.…”

Evaluation of parylene derivatives for use as biomaterials for human astrocyte cell patterning

“…In the absence of piranha acid treatment HEK293 cells showed no preference for parylene-C surfaces over SiO 2 surfaces. Hughes et al has also, however, demonstrated that the piranha acid treatment increases the hydrophilic nature of parylene-C [34].…”