Project description:Stem-cell based cerebral organoids were integrated with 3D-printed perfusable synthetic vasculature. The impact of perfusion was evaluated on month-old cerebral organoids was analyzed by comparing the molecular-level changes in perfused and not-perfused vascularized cerebral organoids
Project description:Human intestinal organoids were grown in a typical 3D matrigel culture environment, or in an alginate gel, then a subset from each condition were xenotransplanted to vascularized and mature in vivo. Epithelium was isolated and epithelial only organoids (enteroids) were then grown from each condition prior to sequencing bulk RNA-sequencing.
Project description:Bulk ATAC-seq was performed on human, chimpanzee, bonobo, and macaque stem cell-derived cerebral organoids. ATAC-seq was performed on day 60 (2 months old) and day 120 (4 months old) cerebral organoids.
Project description:All samples are 50 day old cerebral organoids differentiated from human iPSCs using human the Lancaster protocol (Lancaster et al Nature Protoc, 2014). 4 samples are wild-type (WT) organoids derived from the iPSC line IPSO; 4 samples are Fragile X Syndrome (FX) organoids derived from the iPSC line FX52 in the presence of 15mM HEPES vehicle (Urbach et al, Cell Stem Cell, 2010). 4 samples are Ascorbic Acid treated FX52 cells differentiated into cerebral organoids (FX +AsA). FX +AsA samples were prepared by exposing FX52 iPSCs to 500uM Ascorbic Acid (+15mM HEPES vehicle) for 6 passages (36 days) as iPSCs and then differentiated into cerebral organoids in the presence of Ascorbic Acid throughout the differentiation protocol. By day 50 of the cererbral organoid protocol all organoids were collected for analysis.
Project description:Cerebral organoids co-cultured with patient derived glioma stem cells (GLICOs) are an experimentally tractable research tool useful for investigating the role of the human brain tumor microenvironment in glioblastoma. Here we develop long-term GLICOs, a novel model in which COs are minimally seeded with GSCs and tumor development is monitored over extended durations (ltGLICOs). Single-cell profiling of ltGLICOs revealed an unexpectedly long latency period prior to GSC expansion, and that normal organoid development was unimpaired by the presence of low numbers of GSCs. However, as organoids age they experience chronic hypoxia and oxidative stress which remodels the tumor microenvironment to promote GSC expansion. Receptor-ligand modelling identified astrocytes, which secreted various pro-tumorigenic ligands including FGF1, as the primary cell type for GSC crosstalk and single-cell multi-omic analysis revealed these astrocytes were under the control of ischemic regulatory networks. Functional validation confirmed hypoxia as a driver of pro-tumorigenic astrocytic ligand secretion and that GSC expansion was accelerated by pharmacological induction of oxidative stress. When controlled for genotype, the close association between glioma aggressiveness with patient age has very few proposed biological explanations. Our findings indicate that age-associated increases in cerebral vascular insufficiency and associated regional chronic cerebral hypoxia may contribute to this phenomenon.
Project description:We used cerebral organoids generated from wildtype and CHD8 +/- human ES cells to study the effects of CHD8, one of the top ASD risk genes, on early cortical development. CHD8 +/- hESC were generated using the CRISPR/Cas9 system to create a deletion within the helicase domain. Cerebral organoids were generated according to the protocol from Lancaster et al 2013 with minor modifications.
Project description:LncRNA and mRNA profiling of human iPSC derived cerebral organoids (Propofol treated vs. DMSO control) were determined. LncRNA and mRNA profiling of human iPSC derived cerebral organoids (Propofol treated vs. DMSO control) were determined.
Project description:Single cell ATAC-seq (scATAC-seq) was performed on bonobo induced pluripotent stem cells (iPSC) derived cerebral organoids. scATAC-seq was performed on day 60 (2 months old cerebral organoid) and day 120 (4 months old cerebral organoid).
