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Figure 1.

Sequence of the steps to be performed to prepare MEA substrates before cell plating.

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Figure 1 Expand

Table 1.

Plating parameters of the cultures used in this work.

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Table 1 Expand

Figure 2.

Changes of the number of active channels during cell culture maturation.

Median values include different cell densities for each DIV. Values are normalized over the total number of channels. One-way Friedman test and post-hoc Wilcoxon matched pair test **p<0.01 with respect to DIV 4 and 6.

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Figure 2 Expand

Figure 3.

Median number of active channels at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Values are normalized over the total number of channels. Kruskal-Wallis test and post-hoc Mann-Whitney U test **p<0.01 with respect to cell density of 1800 cells/mm2.

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Figure 3 Expand

Figure 4.

Trend of the number of active channels during cell culture maturation at different cell densities.

Cell culture density equal to 900 cells/mm2 (grey dots), 1800 cells/mm2 (black squares) and 3600 cells/mm2 (white squares). Values are normalized over the total number of channels.

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Figure 4 Expand

Figure 5.

Changes of the mean frequency (Hz) of neuronal networks during cell culture maturation.

Median values include different cell densities for each DIV. One-way Friedman test and post-hoc Wilcoxon matched pair test *p<0.05 with respect to all the other days in culture.

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Figure 5 Expand

Figure 6.

Mean frequency at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Kruskal-Wallis test and post-hoc Mann-Whitney U test *p<0.05 and **p<0.01 with respect to cell density of 1800 cells/mm2.

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Figure 6 Expand

Figure 7.

Mean frequency (in Hz) changes during cell culture maturation at different cell densities.

Cell culture density equal to 900 cells/mm2 (grey dots), 1800 cells/mm2 (black squares) and 3600 cells/mm2 (white squares).

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Figure 7 Expand

Figure 8.

Changes of the burst duration (s) during cell culture maturation.

Median values include different cell densities for each DIV.

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Figure 8 Expand

Figure 9.

Burst length at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Kruskal-Wallis test and post-hoc Mann-Whitney U test #p<0.05 with respect to cell density of 3600 cells/mm2.

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Figure 9 Expand

Figure 10.

Changes of burst duration (s) during culture maturation at different cell densities.

Neuronal culture density equal to 900 cells/mm2 (grey dots), 1800 cells/mm2 (black squares) and 3600 cells/mm2 (white squares).

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Figure 10 Expand

Figure 11.

Changes of the bursting rate (bursts per minute) during cell culture maturation.

Median values include different cell densities for each DIV.

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Figure 11 Expand

Figure 12.

Bursting rate (bursts per minute) at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Kruskal-Wallis test and post-hoc Mann-Whitney U test *p<0.05 with respect to cell density of 1800 cells/mm2.

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Figure 12 Expand

Figure 13.

Bursting rate (bursts/min) during maturation at different cell densities.

Cell culture density equal to 900 cells/mm2 (grey dots), 1800 cells/mm2 (black squares) and 3600 cells/mm2 (white squares).

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Figure 13 Expand

Figure 14.

Changes of network burst length (s) during cell culture maturation.

Median values include different cell densities for each DIV. One-way Friedman test and post-hoc Wilcoxon matched pair test **p<0.01 with respect to all the other days in culture.

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Figure 14 Expand

Figure 15.

Network burst length (s) at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Kruskal-Wallis test and post-hoc Mann-Whitney U test *p<0.05 and **p<0.01 with respect to cell density of 1800 cells/mm2.

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Figure 15 Expand

Figure 16.

Changes of mean duration of NBs (s) during maturation at different cell densities.

Neuronal culture density equal to 900 cells/mm2 (grey dots), 1800 cells/mm2 (black squares) and 3600 cells/mm2 (white squares).

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Figure 16 Expand

Figure 17.

Intra network burst frequency at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Kruskal-Wallis test and post-hoc Mann-Whitney U test #p<0.05 and ##p<0.01 with respect to cell density of 3600 cells/mm2.

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Figure 17 Expand

Figure 18.

Network bursting rate (Hz) during cell culture maturation.

Median values include different cell densities for each DIV. One-way Friedman test and post-hoc Wilcoxon matched pair test *p<0.05 with respect to the first and second weeks (from DIV 4 to DIV 14).

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Figure 18 Expand

Figure 19.

Network bursting rate (Hz) at 900, 1800 and 3600 cells/mm2.

Median values include different DIV for each cell density. Kruskal-Wallis test and post-hoc Mann-Whitney U test +p<0.05 and ++p<0.01 with respect to cell density of 900 cells/mm2.

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Figure 19 Expand

Figure 20.

Network bursting rate (NB/min) during maturation at the three densities under study.

Cell culture density equal to 900 cells/mm2 (grey dots), 1800 cells/mm2 (black squares) and 3600 cells/mm2 (white squares).

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Table 2.

Median (M), 75th percentile (P75) and 25th percentile (P25) for each electrophysiological descriptor analysed and for each cell density (median values include different DIV for each cell density).

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Table 2 Expand