Fig. 1
From: Human cortical spheroids with a high diversity of innately developing brain cell types
Development of neuroectoderm in H9 embryonic stem cell (ESC)-derived human cortical spheroids (hCSs) cultured in the presence of low levels of BMP4 (mild SMAD inhibition) during neurocortical patterning. A Schematic overview of the hCS culturing protocol, depicting crucial proteins and small molecules that induce central nervous system (CNS) cell-type differentiation. The diameters of the generated day-150 (d150) hCSs were between 0.5 and 1.5 mm; scale bar: 1 mm. B Levels of mRNA expression of the stem-cell-specific transcripts NANOG and OCT4, the proliferation-related transcripts MKI67 and TOP2A, and the neural NPC-restricted transcript PAX6 in H9 ECSs and d150 hCSs-1 (batch-1), as determined by quantitative PCR (qPCR) analysis. Independent samples T-test: ***p < 0.001, **p < 0.01. C Levels of mRNA expression of PAX6 in H9 ESCs (d0) and hCSs at various culturing time points of several batches (hCSs2-7). Significances are relative to d0. ANOVA with Dunnett’s T3 post hoc comparisons: *p < 0.05, #p < 0.1. Error bars represent the standard error of the mean. Each data point in (B, C) represents the level of mRNA expression in one spheroid. D Representative immunocytochemistry (ICC) images of neural rosettes (white arrows) containing proliferating KI67-positive cells in d30 hCSs. E Representative ICC-images of PAX6- and SOX9-positive cells in co-stained early (d27) and late (d150) hCSs. The thresholds of the individual color channels in (D, E) were adjusted to better represent the individual signals in the merged image