Project description:Squamocolumnar junction (SCJ) of the uterine cervix is a target of human papilloma virus (HPV) infection and thereby a putative cell-of-origin for uterine cervical cancer. However at present, in vitro models that accurately phenocopy its homeostasis is lacking. In this study, we aimed at establishing organoids of SCJ cells and identifying its characteristics. Samples were collected from the uterine cervix of 4 cases who underwent hysterectomy at our department, and processed to initiate organoid culture. Organoids were obtained from 3 samples among them, and they were analyzed both histologically and by transcriptome analysis.
Project description:Organoid culture is important for maintenance of epithelial cell characteristics, stemness, and tumorigenic activity of biliary tract cancer initiating cells. To investigate whether organoid culture maintain cancer stem cell properties of biliary tract cancer initiating cells, we compared the gene expression changes between organoid culture and adherent culture.
Project description:An organoid culture system can better recapitulate the cellular structure, function, and interaction between cells and the extracellular matrix (ECM) than the traditional two-dimensional (2D) culture system. We here constructed a condylar cartilage organoid and utilized it to explore the regulatory role of primary cilia at the organoid level. RNA sequencing unveiled the differences of transcriptomics between the condylar cartilage organoid and 2D culture chondrocytes.
Project description:Organotypic in vitro culture is useful to model mammalian disease in numerous tissues. Normal epithelial differentiation and carcinogenesis both undergo in vivo regulation by stroma, but current culture methods exclude stroma. To mimic this in vivo environment, we developed and characterized a human 3D prostate organoid co-culture model that incorporates prostate stroma. Primary prostate stromal cells supported increased organoid formation and expressed growth factors and WNT-related genes involved in epithelial differentiation. Organoid branching occurred distal to physical contact with stromal cells, demonstrating non-random branching. Tumoroids derived from primary prostate cancer maintained differential expression of the prostate cancer marker AMACR only in the presence of stroma. Stroma-induced phenotypes were similar in all patients examined, yet maintained inter-patient heterogeneity in the degree of response. Addition of stroma to in vitro organoid culture recapitulated the in vivo microenvironment by inducing organization of benign organoids into branching structures and preserving prostate cancer phenotypes.
Project description:Stem-cell-derived epithelial organoids are routinely used for the biological and biomedical modelling of tissues. However, the complexity, lack of standardization and quality control of stem cell culture in solid extracellular matrices hampers the routine use of the organoids at industrial scale. Here, we report the fabrication of microengineered cell-culture devices and scalable and automated methods for the suspension culture and real-time analysis of thousands of individual gastrointestinal organoids trapped in microcavity arrays within a polymer-hydrogel substrate. The absence of a solid matrix significantly reduces organoid heterogeneity, as we show for mouse and human gastrointestinal organoids. We used the devices to screen for anticancer drug candidates with patient-derived colorectal cancer organoids, and high-content image-based phenotypic analyses to reveal insights into drug-action mechanisms. The scalable organoid-culture technology should facilitate the use of organoids in drug development and diagnostics.
Project description:Transcriptomic profiles of 6 commercially-available human patient-derived gastrointestinal organoid lines were obtained and compared to transcriptomic profile of a commercially available human iPSC-induced colon organoid line. Transcriptomic profile of iPSC-derived human colon organoid line was compared after culture in either Corning growth-factor-reduced Matrigel (Corning 356231) or MilliporeSigma growth-factor-reduced ECMGel (E6909)
