Patterning and detailed study of human hNT astrocytes on parylene-C/silicon dioxide substrates to the single cell level

Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining

“…For single pulse high power ablation the 20um wide strips provided a 28% cellular coverage and 12% nuclear coverage and a 5.6:1 contrast between the cells on the parylene-C and those on the SiO 2 substrate. These results are comparable to our previously published hNT astrocyte patterning on photolithographic produced parylene-C/SiO2 substrates [8] which provided an astrocyte cell process coverage of 40% with 12% nuclear coverage for the same parylene-C strip dimensions.…”

“…The percentage of the substrate covered by cells was measured for three regions, 'on the parylene-C', 'adjacent to the parylene-C' and 'off the parylene-C,' as shown in Figure 1. A more detailed description of the image pre-processing steps can be found in Unsworth et al [8].…”

Enrichment of differentiated hNT neurons and subsequent analysis using flow-cytometry and xCELLigence sensing

“…methods involving transcriptomics, or proteomics), the use of highly pure or enriched neuronal cultures is desirable. We have modified our hNT differentiation protocol (Raos et al, 2013;Unsworth et al, 2011), to better facilitate enrichment of the neuronal yield. This adapted method preferentially selects the immature-astrocyte like cells by rapid adhesion to plastic (the pre-astrocytic cells have a strong preference for Nunc), and leaves the majority of the neurons in suspension.…”

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

“…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).…”

“…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.…”