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Patterning of excitable cells on multi-electrode arrays
Slavík, Jan ; doc. RNDr. Petr. Skládal, CSc. (oponent) ; Průcha,, Jaroslav (oponent) ; Hubálek, Jaromír (vedoucí práce)
The work deals with the alignment of excitable cells on multielectrode arrays. First, the alignment of excitable cells was analyzed. Embryonic neurons from rat hippocampus and HL-1 cells, which are derived from the AT-1 line of tumor mouse atrial cardiomyocytes, were used. Alignment was tested on ridged surfaces and on surfaces with materials with different cellular affinities. It was demonstrated, that both neurons and HL-1 cells aligned in direction of ridges on ridges surface, but only neurons aligned on surface with different chemical affinity. Further, own multielectrode arrays were made, the HL-1 cells were cultured on the multielectrode arrays, and the action potentials of the HL-1 cells were measured and analyzed. The aim was to prove that it is possible to measure the action potential on the fabricated multi-electrode arrays. A special multielectrode array with a uniform surface was made to align cells on a multielectrode array. This multielectrode array is called a planar multielectrode array. The planar multielectrode arrays were made by a special fabrication process. The layers of the planar multielectrode array were deposited on the sacrificial substrate in the reverse order. The sacrificial substrate for deposition was silicon wafer on which was deposited another sacrificial layer from gold. The upper insulating layer of planar multielectrode array was deposited first and the lowest substrate layer was deposited last. Then the planar multielectrode array with the sacrificial gold layer was peeled off from silicon due to the low adhesion of gold to silicon and planar multielectrode array turned upside down. The sacrificial gold layer was removed by a wet etchant and planar multielectrode array was finished. On a planar multielectrode array, the HL-1 cells were patterned into strips by a chemical method using a combination of microprinting of an adhesive agent and subsequent coating by an anti-adhesive agent of not microprinted areas. The electrophysiological properties of aligned HL-1 cells were measured using the planar multielectrode array. By this experiment, it was introduced fabrication technology for fabrication of planar multielectrode arrays and the planar multielectrode array was successfully tested for HL-1 cells alignment on its surface by combination of microprinting of an adhesive agent and anti-adhesive agent coating.

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