Project description:Microarray analysis was performed on P0 SFTPC+ cells, P0 SFTPC- cells and CPMhigh progenitor cells derived form human induced pluripotent stem cells (B2-3 SFTPC-GFP reporter hiPSCs), respectively.
Project description:We developed LB cells overexpressing ABCA3, SFTPB, SFTPC, and SFTPD in A549 cells. Using LB cells in high content screening, we identified HPbCD. HPbCD improved amiodarone-induced LB abnormality in iPS cell-derived alveolar organoids.
Project description:We utilized patient-derived induced pluripotent stem cells (iPSCs) to generate 3D cerebral organoids to model neuropathology of Scz during this critical period. We discovered that Scz organoids exhibited ventricular neuropathology resulting in altered progenitor survival and disrupted neurogenesis. cz organoids principally differed not in their proteomic diversity, but specifically in their total quantity of disease and neurodevelopmental factors at the molecular level. Provides unique insights into the proteome landscape of schizophrenia in patient-derived cerebral organoids
Project description:We stepwisely induced SFTPC+ cells from hiPSCs via CPM-high progenitor cells with or without coculturing human fetal lung fibroblasts. Single-cell RNA-seq of CPM-high progenitor and hiPSC-derived SFTPC+ cells demonstrated their differentiation process and cellular heterogeneity.
Project description:Alveolar type 2 (AT2) organoids, derived from induced pluripotent stem cells, were stimulated with a fibrosis cocktail containing the pro-fibrotic and inflammatory cytokines TGF-β (5 ng/ml), TNF-α (10 ng/ml), PDGF-AB (10 ng/ml) and LPA (5 μM) or control cocktail containing diluents for 72 h.
Project description:Organoid derived from human induced pluripotent stem cells (hiPSC) is potentially applicable for regenerative medicine. However, the applications have been hampered by limited organoid size and function as a consequence of a lack of progenitor expansion. Here, we report the recapitulation of the in vivo progenitor expansion in hiPSC-liver organoid based on the analysis of mouse development. Visualization of blood perfusion and oxygen levels in mouse embryos revealed a transient hypoxic environment despite blood flow while hepatoblast expansion. During this specific stage, the placenta was seen to express various growth factors. Human and mouse placenta-liver interaction analysis identified various placenta-derived factors. Among them, IL1α efficiently induced the growth in hiPSC-liver organoids as well as mouse fetal livers following progenitor expansion under hypoxia. Subsequent oxygenation demonstrated that expanded progenitors by IL1α contributed to hiPSC-liver organoid size and function. Taken together, treatment of placenta-derived factor under hypoxia is a crucial organoid culture technique that efficiently induces progenitor expansion.
Project description:We found that hepatocytes isolated from adult mouse livers can dedifferentiate into progenitor-like cells and exhibit a hybrid epithelial/mesenchymal phenotype in monolayer culture. dediHeps can re-differentiate into mature hepatocytes by forming aggregates and fetal intestinal progenitor cells (FIPCs) by forming organoids in three-dimensional culture. We conducted RNA-seq analyses to investigate the characteristincs of hepatocytes, dediHeps, dediHep aggregates, dediHep derived spherical organoids, dediHep derived budding organoids, FIPC derived spherical organoids, intestinal stem cell (ISC) derived budding organoids, and CLiP cells (Katsuda, 2016).
Project description:To address the need for human alveolar epithelial cell (AEC)-derived lines to more suitably model distal lung diseases, we have generated and characterized novel immortalized cell lines derived from human AECs. We used a combination of the ROCK inhibitor, Y-27632, and lentiviral transduction of SV40 Large T antigen of previously cryopreserved isolated human alveolar epithelial type 2 (AT2) cells to generate immortalized AECs. These AEC lines proliferate well on standard tissue culture dishes forming an epithelial monolayer and express lung progenitor markers SOX9 and SOX2. When grown in 3D culture with lung fibroblasts, the cells form NKX2-1+ organoids expressing more mature alveolar lung markers, AQP5 and GPRC5A. Single cell RNA-sequencing of one AEC line comparing cells in 2D versus 3D revealed increased cellular heterogeneity and an induction of cytokine and lipoprotein signaling in 3D culture, reflecting interactions with the microenvironment during organoid formation. Taken together, these data show our novel progenitor-like AEC lines retain a genetic and structural memory of their alveolar cell lineage despite long-term expansion, providing a valuable new system to model the distal lung in vitro.
