Fig. 1

3D cultures generated from sclerotic hippocampi of patients with mTLE replicate components of the in-vivo stem cell niche. A Schematic representation of air-liquid interface 3D culture generation. Tissue sections for immunohistochemical staining were obtained by cryosectioning. B Early neuronal progenitors (Sox2⁺) are present in our 3D cultures (Scale bar 100 μm). C Both nestin⁺ GFAP⁺ type 1 cells and nestin⁺ GFAP⁻ type 2a cells, as well as nestin⁻ GFAP⁺ astrocytes are present (scale bar 100μm). D The 3D cultures also contain many immature (NeuroD1⁺) neurons, some of which have been formed in culture, as indicated by EdU incorporation (Scale bar 100 μm). E The NeuroD1⁺ neurons are dentate gyrus derived, as shown by expression of the developmentally specific granule cell neuron marker Prox1 (scale bar 100 μm). F βIII tubulin staining reveals the complex morphology of neurons in 3D culture (Scale bar 100 μm). G The presence of recently generated mature post-mitotic neurons is indicated by NeuN and BrdU co-immunoreactivity, indicating generation in culture (scale bar 25 μm). H Small numbers of IB4⁺ microglia are present (scale bar 25 μm). I Many of the remaining cells are PECAM⁺ endothelial cells. scale bar 10μ (scale bar 100 μm)