Effect of Cell-Adhesive Nanofibril on 3D spheroid hepatic differentiation-related gene expression
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ABSTRACT: To investigate the effect of Cell-Adhesive Nanofibril on the hepatic differentiation process of 3D iPSC spheroids, we established hepatic organoids that underwent a stepwise hepatic differentiation process with or without Cell-Adhesive Nanofibril.
Project description:We utilize bulk RNA-seq to profile the mRNA expression in day 3 and day 14 post-implantation of SD rat abdminal wall treated with adhesive or non-adhesive implants
Project description:Tumor angiogenesis is regarded as a promising target for limiting cancer progression because tumor-associated vasculature supplies blood and provides a path for metastasis. Thus, in vitro recapitulation of vascularized tumors is critical to understand the pathology of cancer and identify the mechanisms by which tumor cells proliferate, metastasize, and respond to drugs. In this study, we microengineered a vascularized tumor spheroid (VTS) model to reproduce the pathological features of solid tumors. We first generated tumor cell-EC hybrid spheroids with self-assembled intratumoral vessels, which enhanced the uniformity of the spheroids and peritumoral angiogenic capacity compared to spheroids comprised only with cancer cells. Notably, the hybrid spheroid also exhibited expression profiles associated with aggressive behavior. The blood vessels sprouting around the hybrid spheroids on the VTS chip were interconnected with intratumoral vessels and displayed the distinctive characteristics of leaky tumor vessels. With the VTS chip showing a progressive tumor phenotype, we validated the suppressive effects of axitinib on tumor growth and angiogenesis, which depended on exposure dose and time, highlighting the significance of tumor vascularization to predict the efficacy of anticancer drugs. Ultimately, we effectively induced both lymphangiogenesis and angiogenesis around the tumor spheroid by promoting interstitial flow. Thus, our VTS model is a valuable platform with which to investigate the interactions between tumor microenvironments and explore therapeutic strategies in cancer.
Project description:Remarkable advancements in protocol development have been achieved to manufacture insulin-secreting islets from human pluripotent stem cells. Distinct from current approaches, we devised a tunable strategy to generate islet spheroids enriched for major islet cell types by incorporating PDX1+ cell budding morphogenesis into the differentiation process of staged transcriptional programming. In this process that appears to mimic normal islet morphogenesis, the differentiating islet spheroids self-organize with endocrine cells that are intermingled or arranged in a core-mantle architecture, accompanied with functional heterogeneity. Through in vitro modelling of human pancreas development, we illustrate the importance of PDX1 in eliciting cell budding morphogenesis, and the requirement for EphB3/4 signaling. We show how RFX6 deficiency affects pancreatic patterning and uncover an expected role of RFX6 in early pancreas morphology. The tunable differentiation system and stem cell-derived islet models described in this work may facilitate addressing fundamental questions in islet biology and probing human pancreas diseases.
Project description:Microarray based mRNA profiling was used to charactarize and compare the gene expression in cells grown as monolayer or spheroids. Microarray based mRNA profiling was used to charactarize and compare the gene expression in cells grown as monolayer or spheroids.
Project description:Tumor microenvironments present significant barriers to anti-tumor agents. Molecules involved in multicellular tumor microenvironments, however, are difficult to study ex vivo. Here, we generated a matrix-free tumor spheroid model using the NCI-H226 mesothelioma cell line and compared the gene expression profiles of spheroids and monolayers using microarray analysis. Microarray analysis revealed that 142 probe sets were differentially expressed between tumor spheroids and monolayers. Gene ontology analysis revealed that upregulated genes were primarily related to immune response, wound response, lymphocyte stimulation and response to cytokine stimulation, whereas downregulated genes were primarily associated with apoptosis. Among the 142 genes, 27 are located in the membrane and related to biologic processes of cellular movement, cell-to-cell signaling, cellular growth and proliferation and morphology. Western blot analysis validated elevation of MMP2, BAFF/BLyS/TNFSF13B, RANTES/CCL5 and TNFAIP6/TSG-6 protein expression in spheroids as compared to monolayers. Thus, we have reported the first large scale comparison of the transcriptional profiles using an ex vivo matrix-free spheroid model to identify genes specific to the three-dimensional biological structure of tumors. The method described here can be used for gene expression profiling of tumors other than mesothelioma. We generated a matrix-free tumor spheroid model using the NCI-H226 mesothelioma cell line and compared the gene expression profiles of spheroids and monolayers using microarray analysis. Microarray analysis experiments were performed in triplicates. A total of six samples (three for monolayers and three for spheroids) were analyzed.
Project description:To investigate epigenetic dynamics in a 3D liver spheroid model, liver spheroids were treated with sodium butyrate for 3 days and allowed to recover for 1 week. We then performed gene expression profiling analysis using data obtained from RNA-seq of spheroids at 4 time points
Project description:Recently, it has been proposed to employ three-dimensional cultivation of cell in spheroids that contributes more proper physiological conditions in matrix extracellular space architecture and cell morphology accompanied by appropriate intracellular biomechanical organization of cytoskeleton and energy generation apparatus adaptation. In this research we presented the data of proteome survey for cells growing in monolayer and in spheroids that were induced in myogenic way of differentiation and reached terminal stage in seven days. Semi-quantitative profiling of the dynamic changes in proteins abundance depicted strict advantage of spheroid manner cultivation in contrast to monolayer. Cells cultured in spheroid were significantly enriched in proteins of mitochondria biogenesis, respiratory chain proteins, extracellular proteins and cytoskeleton. Most of signal transducers involved in active proliferation and early stages of differentiation were inhibited in spheroid cultures unlike monolayer. The obtained data were broadly consistent with known molecular mechanisms of myogenesis in transcripts level and reflected strong advantage of spheroid cultivation conditions.
Project description:Microarray based mRNA profiling was used to charactarize and compare the gene expression in cells grown as monolayer or spheroids. Keywords: Expression profiling by array Microarray based mRNA profiling was used to charactarize and compare the gene expression in cells grown as monolayer or spheroids.