Generation of functional hippocampal neurons from self-organizing human embryonic stem cell-derived dorsomedial telencephalic tissue

Abstract

The developing dorsomedial telencephalon includes the medial pallium, which goes on to form the hippocampus. Generating a reliable source of human hippocampal tissue is an important step for cell-based research into hippocampus-related diseases. Here we show the generation of functional hippocampal granule- and pyramidal-like neurons from self-organizing dorsomedial telencephalic tissue using human embryonic stem cells (hESCs). First, we develop a hESC culture method that utilizes bone morphogenetic protein (BMP) and Wnt signalling to induce choroid plexus, the most dorsomedial portion of the telencephalon. Then, we find that titrating BMP and Wnt exposure allowed the self-organization of medial pallium tissues. Following long-term dissociation culture, these dorsomedial telencephalic tissues give rise to Zbtb20+/Prox1+ granule neurons and Zbtb20+/KA1+ pyramidal neurons, both of which were electrically functional with network formation. Thus, we have developed an in vitro model that recapitulates human hippocampus development, allowing the generation of functional hippocampal granule- and pyramidal-like neurons.

Three-dimensional induction of dorsal, intermediate and ventral spinal cord tissues from human pluripotent stem cells

“…hESCs were maintained and cultured as previously described (Sakaguchi et al, 2015). In brief, hESCs were maintained on a feeder layer of mouse embryonic fibroblasts inactivated by mitomycin C treatment in DMEM/F12 (Wako) supplemented with 20% knockout serum replacement (KSR, Invitrogen), 2 mM glutamine, 0.1 mM nonessential amino acids (Invitrogen), 5 ng ml −1 recombinant human basic fibroblast growth factor (bFGF) (Wako) and 0.1 mM 2-mercaptoethanol under 2% CO 2 .…”

Modeling Development and Disease with Organoids

“…A final protocol developed by Sasai and coworkers involved the in vitro generation of a reliable source of hippocampal tissue from human ES cells (Sakaguchi et al, 2015). SFEBq served once again as the starting material.…”

Using brain organoids to understand Zika virus-induced microcephaly

“…The cerebral organoid offers an opportunity to model whole brain development and interactions among different brain regions. Brain region-specific organoids, by contrast, use patterning factors to induce the differentiation of hPSCs into specific lineages, such as cerebral cortex (Kadoshima et al, 2013;Mariani et al, 2015;Pasça et al, 2015;Qian et al, 2016), midbrain (Jo et al, 2016;Qian et al, 2016) and hypothalamus (Merkle et al, 2015;Qian et al, 2016;Sakaguchi et al, 2015;Wang et al, 2015). Compared with cerebral organoids, brain region-specific organoids model individual brain regions of interest and generally result in more uniform and reproducible tissue, providing a platform for quantitative characterization.…”