Fig 2: The neuroepithelial buds and cerebral organoids recapitulated early stages of human fetal brain development.(A) Schematic diagram depicting the effect of HSV-1-infection on the neuroepithelial buds and brain organoids in the areas of modelling the pathological features of developing human fetal brain, including injured neurogenesis, impaired neuronal differentiation, abnormal microglial activation, and dysregulated brain regionalization. (B) Schematic diagram of the neuroepithelial bud and cerebral organoid method and timing. Scale bars: 50µm. (C) The immunofluorescence staining for PAX6, TUJ, Nestin, and SOX2 in the neuroepithelial buds at the day 18. Scale bars: 25 µm. (D) Expressions of the specific brain regions markers (forebrain, PAX6; hindbrain ISL1) in brain organoids at the day 18; the cerebral organoids derived from hiPSCs at the day 45 showed the diverse neuron subtypes, including the mature neurons (MAP2), the astrocyte marker (GFAP), and microglia markers (Iba1). Scale bars: 25 µm. The images were taken Images were captured on a Leica TCS SP8 STED confocal microscope.

Fig 3: Spinal inflammatory response, number of Nissl bodies and activation of glial cells of sham and cancer induced bone pain (CIBP) rats. (A) Inflammatory cells infiltration of lumbar spinal dorsal horn from sham and CIBP rats were examined by Haematoxylin and eosin staining. Scale bar: 50 µm. (B) Nissl bodies in spinal dorsal horn from sham and CIBP rats were detected by Nissl staining. Scale bar: 50 µm. (C) Quantitative analysis of inflammation score in (A). *p < 0.05 vs. sham group. (D) Quantitative analysis of numbers of Nissl positive cells. *p < 0.05 vs. sham group. (E and F) Representative images of Iba1 and GFAP immunostaining on sections of spinal cords from sham and CIBP rats. (G and H) Quantitative analysis of Iba1 (E) and GFAP (F) fluorescence intensity in spinal dorsal horns from sham and CIBP rats. Scale bars: first line, 200 µm; second line, 20 µm. Data were showed as the mean ± SD. *p < 0.05 vs. sham group. Data represents five independent experiments