Fig. 4: Generation and functional characterization of hASA.
From: Human assembloid model of the ascending neural sensory pathway
a, Generation of hASA. b,c, Bright-field (b) and fluorescent image (c) of hASA. d,e, Calcium imaging (d) and traces (e) after αβ-MeATP exposure. f, Responses following αβ-MeATP exposure, compared to null value (0), in organoids (hCO, n = 83 cells; hDiO, n = 52 cells; hdSpO, n = 84 cells; hSeO, n = 131 cells from three to four organoids; two differentiations) and assembloids (hCO, n = 91 cells; hDiO, n = 116 cells; hdSpO, n = 98 cells; hSeO, n = 95 cells from six assembloids; one differentiation) on daf 74–78. Wilcoxon signed-rank test; ****P < 0.0001. g,h, Glutamate uncaging after ultraviolet application to hSeO (g) and ΔF/Fbase traces (h). i, Response before and after ultraviolet exposure (hCO, n = 113 cells; hDiO, n = 157 cells; hdSpO, n = 159 cells; hSeO, n = 109 cells; five assembloids; one differentiation; Wilcoxon matched-pair signed-rank test; ****P < 0.0001) on daf 74–99. j, Calcium imaging. k,l, jGCaMP8s expression, spontaneous activity, SCA correlograms (hSeO–hCO pair) and peak percentage correlations in organoids (k) and assembloids (l). Asterisks; synchronous events. m, Correlation values in organoids (O; hCO, n = 96 cells; hDiO, n = 61 cells; hdSpO, n = 63 cells; hSeO, n = 47 cells; seven organoids; three differentiations) and assembloids (A; hCO, n = 208 cells; hDiO, n = 157 cells; hdSpO, n = 156 cells; hSeO, n = 110 cells; ten assembloids; three differentiations) on daf 74–131. Mann–Whitney test; ****P < 0.0001. n,o, Four-part extracellular recordings (n) and traces (o). Asterisks; synchronous events. p, Representative cross-correlograms. q, Co-activation index for hSeO–hdSpO, hSeO–hDiO and hSeO–hCO pairs (n = 223, 180 and 222 channels, respectively; seven to eight assembloids; four differentiations; Wilcoxon matched-pair signed-rank test; ****P < 0.0001) on daf 80–99. Data: mean ± s.e.m. Small dots, cells or channels; large dots, assembloids. Scale bars, 1 mm (b), 500 μm (c), 400 μm (k,l). Veh., vehicle. Credits: a,d,g,j adapted from ref. 48, Springer Nature America, Inc.; n adapted with permission from ref. 11, Elsevier.
