Journal: Cell Reports Methods

Article Title: Profiling human iPSC-derived sensory neurons for analgesic drug screening using a multi-electrode array

doi: 10.1016/j.crmeth.2025.101051

Figure Lengend Snippet: hiPSC nociceptor growth, neural activity, Z′, and robust Z′ factors on a 48-well MEA plate hiPSC nociceptors were cultured on a 48-well microelectrode array (MEA) plate for 4 weeks. (A) Representative micrographs of hiPSC nociceptors spot seeded at different cell densities (2,000, 35,000, and 65,000 cells/well) and two time points (day 2 and day 28). These micrographs were imaged at 100× magnification. (B–E) Across the 4 weeks of culture, cells were evaluated for (B) total spike count per well; (C) mean firing rate per well; (D) impedance, a measure of cell viability of hiPSC nociceptors; and (E) active electrode yield (AEY), indicating the percentage of electrodes successfully recording neural activity. At the 28 day time point, cells were challenged with inhibitors, and the Z′ and robust Z′ factors were calculated to measure the quality of the assay. (F and G) Z′ (F) and robust Z′ (G) factors were calculated at different densities, showing high quality with the robust Z′ factor at several seeding densities. (H) Scatterplots comparing well total spike count data before (blue) and 2 h post treatment with 500 μM lidocaine (pink) for a representative density of 35,000 cells/well. (I) Normal quantile-quantile (QQ) plot assessing the normality of data distribution before and after lidocaine treatment (five wells for each density). Next, two independent experiments were conducted to evaluate the Z′ and robust Z′ of the assay to three neuronal inhibitors (lidocaine, tetrodotoxin [TTX], and huwentoxin [HWTX]) for cells at 28 days in culture at 35,000 cells/well. (J and K) Z′ factor (J) and the robust Z′ values (K) for different inhibitors in experiment 1 (12 wells for each inhibitor). (L and M) Z′ factor (L) and the robust Z′ values (M) for different blockers in experiment 2 using 35,000/well (12 wells for each inhibitor). 0.5 dotted lines represent the Z′ threshold for a “good” quality assay. Data from these experiments represent three distinct sets of experiments, including two confirmatory experiments as shown. Data from (B)–(E) are presented as mean ± SEM (standard error of the mean).

Article Snippet: The goal was to leverage the human cellular heterogeneity and neuronal firing of the hiPSC nociceptors to build a strong high-content screening (HCS) assay using a multi-well microelectrode array (MEA) platform for pain therapeutics.

Techniques: Activity Assay, Cell Culture, Microelectrode Array