Journal: Scientific Reports

Article Title: Chronic Electrical Stimulation Promotes the Excitability and Plasticity of ESC-derived Neurons following Glutamate-induced Inhibition In vitro

doi: 10.1038/s41598-018-29069-3

Figure Lengend Snippet: ESC-derived neurons and glia form functional neural network on MEA. ( A ) Experimental schedule for neural stem cell seeding and culture. For the control group, recordings were performed from week1 to week 3. ( B ) Representative micro-electrode array (MEA) recording setup. Schematic of the bottom of the well with 4 reference electrodes and 8 × 8 electrode array (left). Top left quadrant of a 64-channel MEA plate, 2 weeks after stem cells seeding (right; Scale = 100 μm). ( C ) Scatter distribution of the spike width (µsec; x-axis) against the peak-to-valley amplitude (µVolt; y-axis) for individual unit recorded from control condition at week 3. The distribution (cell count per bin) is displayed as a projection of each axis. The dashed black line represents the average of spike width (vertical) and peak-to-valley amplitude (horizontal). ( D ) Representative average spike wave forms obtained from 3 electrodes at week 3 from the control group. Data is shown as mean and s.e.m. ( E ) Raster plot of one well recorded at week 3 (top panel). The population instantaneous firing rate (left y-axis in green; wMFR, spikes/bin/electrode) and the percentage of active electrodes per bin (right y-axis, in red; active electrode in %). The vertical dashed and dotted lines indicate the start and stop of a detected population burst, respectively. ( F ) Heat map showing the evolution of the average activity in a control well from week 2 to week 3.

Article Snippet: Briefly, each well of 12 well/64 electrode per well containing MEA plates (Axion Biosystems, GA), or glass bottom petri dishes (Cellvis, CA) were coated with polyethylenimine (PEI) (Sigma-Aldrich, MO) and incubated for 1 h at 37 °C and 5% CO 2 , then washed with deionized water and allowed to air dry overnight.

Techniques: Derivative Assay, Functional Assay, Control, Cell Counting, Activity Assay

Journal: Scientific Reports

Article Title: Chronic Electrical Stimulation Promotes the Excitability and Plasticity of ESC-derived Neurons following Glutamate-induced Inhibition In vitro

doi: 10.1038/s41598-018-29069-3

Figure Lengend Snippet: Acute L-glutamate treatment impaired network activity and synchrony immediately and over a week post-administration. ( A ) Experimental schedule for ESC-derived neuron seeding, culture and L-glutamate treatment. In the treatment group, 100 µM of L-glutamate was administered at day14 for 20 min then washed out. MEA recordings were performed from week 1 to week 3. ( B ) Scatter distribution of the mean burst firing rate (mBR; Burst event/min; x-axis) against the mean firing rate (mFR; log of spike/sec; y-axis) for individual unit recorded from CTR (black filled circles) and L-glut (black open circle) groups. Data is shown for week 2 (left panel) and week 3 (right panel). The normalized distribution CTR (black line) and L-glut (gray line) groups are displayed as a projection of each axis. The dashed black line represents the average for the CTR group. The dotted black line represents the average for the L-glut group. mFR: mean firing rate for individual electrodes; mBR: mean bursting rate for individual electrodes. ( C ) ESC-derived neurons after L-glutamate treatment (L-glut; 100 µM for 20 min at day 14) at week 2 (top panels) and week 3 (bottom panels). From left to right panels are shown SOX-1, βIII-Tubulin and merged image of both fluorescent markers. Scale = 200 μm. ( D ) Change at week 2 and week 3 (expressed as a Z-score from week1 baseline) of the number of bursting cells (left panel) and weighted mean firing rate (wMFR; right panel). For each group, the number of wells with significantly increasing change over the total number of wells recorded is shown above the scatter plot. wMFR: weighted mean population firing rate. ( E ) Change in network synchrony at week 2 and week 3 (expressed as a Z-score from week1 baseline) for event synchronization (left panel), cross-correlation peak (middle panel) and mean network burst firing rate (mBFR; right panel). For each group, the number of wells with significantly increasing change over the total number of wells recorded is shown above the scatter plot. For post-hoc two-sample test *, ** and *** indicate p < 0.05, p < 0.01 and p < 0.001, respectively. mBFR: mean population burst firing rate.

Article Snippet: Briefly, each well of 12 well/64 electrode per well containing MEA plates (Axion Biosystems, GA), or glass bottom petri dishes (Cellvis, CA) were coated with polyethylenimine (PEI) (Sigma-Aldrich, MO) and incubated for 1 h at 37 °C and 5% CO 2 , then washed with deionized water and allowed to air dry overnight.

Techniques: Activity Assay, Derivative Assay

Journal: Scientific Reports

Article Title: Chronic Electrical Stimulation Promotes the Excitability and Plasticity of ESC-derived Neurons following Glutamate-induced Inhibition In vitro

doi: 10.1038/s41598-018-29069-3

Figure Lengend Snippet: DCS and LFS stimulation enhanced the maturation in excitability and neural synchrony following acute L-glutamate treatment. ( A ) Experimental schedule for ESC-derived neuron seeding, culture and treatment. L-glutamate (100 µM) was administered at day14 for 20 min then washed out. LFS was administered from day 15 to day 19 using 10 µA at 0.1 Hz, 15 min/day. DCS was administered in two phases: day14 a cathodal stimulation was performed for 15 min; from day 15 to day 19, anodal stimulation was administered for 15 min per day. Recording was performed on day 14 (week 2) and day 21 (week 3). ( B ) Schematic set up for electrical stimulation on MEA plate. LFS stimulation (10 µA, 0.1 Hz, 15 min/day) was delivered through the MEA electrode using the Maestro system. Four stainless screws were positioned above the MEA cultured neurons and were used to deliver a controlled current (DCS: single-time 10 µA monophonic cathodal 15 min and daily 10 µA monophonic anodal current, 15 min/day) using a custom battery-powered system. ( C ) Change at week 3 (expressed as a Z-score from week1 baseline) of the number of bursting cells (left panel) and wMFR (right panel). For post-hoc multiple comparison using Dunn-Sidak correction *, ** and *** indicate p < 0.05, p < 0.01 and p < 0.001, respectively. wMFR: weighted mean population firing rate. ( D ) Change at week 3 (expressed as a Z-score from week1 baseline) of the event synchronization (left panel), cross-correlation peak (middle panel) and mBFR (right panel). For post-hoc multiple comparison using Dunn-Sidak correction *, ** and *** indicate p < 0.05, p < 0.01 and p < 0.001, respectively. mBFR: mean population burst firing rate.

Article Snippet: Briefly, each well of 12 well/64 electrode per well containing MEA plates (Axion Biosystems, GA), or glass bottom petri dishes (Cellvis, CA) were coated with polyethylenimine (PEI) (Sigma-Aldrich, MO) and incubated for 1 h at 37 °C and 5% CO 2 , then washed with deionized water and allowed to air dry overnight.

Techniques: Derivative Assay, Cell Culture, Battery, Comparison

Journal: bioRxiv

Article Title: Multielectrode array characterization of human induced pluripotent stem cell derived neurons in co-culture with primary human astrocytes

doi: 10.1101/2024.03.04.583341

Figure Lengend Snippet: 24-well quantification of (A) Mean firing rate (Hz), (B) Number of active electrodes, (C) Network burst frequency (Hz), (D) Network burst duration (s), (E) Network interburst interval coefficient of variation, and (F) Synchrony index on DIV 11, 17, 26, and 33. (G) Representative raster plots. Statistical analysis was performed with repeated measures ANOVA and Bonferroni corrections Error bar: SEM. N=24 independent wells. p-value < 0.05 *, p-value < 0.01 **, p-value < 0.001 ***

Article Snippet: Multielectrode array 24-well plates containing 16 electrodes per well were obtained from Axion Biosystems (CytoView MEA plate).

Techniques:

Journal: bioRxiv

Article Title: Multielectrode array characterization of human induced pluripotent stem cell derived neurons in co-culture with primary human astrocytes

doi: 10.1101/2024.03.04.583341

Figure Lengend Snippet: Characterized by (A) Mean firing rate (Hz) (B) Number of active electrodes (C) Network burst frequency (Hz) (D) Average network burst duration (s) (E) Network interburst interval coefficient of variation (F) Synchrony index. Statistical analysis was performed with an independent t-test. Error bar: SEM. N=24 independent wells per plate. p-value < 0.05 *, p-value < 0.01 **, p-value < 0.001 ***

Article Snippet: Multielectrode array 24-well plates containing 16 electrodes per well were obtained from Axion Biosystems (CytoView MEA plate).

Techniques:

Journal: bioRxiv

Article Title: Multielectrode array characterization of human induced pluripotent stem cell derived neurons in co-culture with primary human astrocytes

doi: 10.1101/2024.03.04.583341

Figure Lengend Snippet: (A) Representative raster plots and histograms of relative population firing rates for baseline, 24 hours after treatment, and after 2 washouts. (B) Mean firing rate (Hz) (C) Number of active electrodes (D) Network burst frequency (Hz) (E) Network burst percentage (%) (F) Synchrony Index (G) Network Interburst Interval Coefficient of Variation. Statistical analysis was performed with repeated measures ANOVA and Bonferroni corrections. Error bar: SEM. N=4 independent wells. p-value < 0.05 *, p-value < 0.01 **, p-value < 0.001 ***

Article Snippet: Multielectrode array 24-well plates containing 16 electrodes per well were obtained from Axion Biosystems (CytoView MEA plate).

Techniques: