Project description:In order to study the role of piR_019914 in C28/I2 cells, we established piR_019914_mimics to overexpress piR_019914 in C28/I2 cells We then performed gene expression profiling analysis using data obtained from RNA-seq of piR_019914 overexpression C28/I2 cells and mimics_NC C28/I2 cells.
Project description:3-D suspension culture differs considerably from monolayer culture in regards to cell architecture, cell to cell contact, and cell to matrix interactions. These factors play a regulatory role in epithelial stem cell homeostasis and influence epidermal stem cells on their decision to remain quiescent or to activate self-renewal. We used microarrays to determine the global programme of gene expression underlying spheroid ring formation and identified distinct gene programs involved in this process.
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:Anchorage-independent spheroid cells of HCC possess stemness properties. Huh7 cells are cultured in ultra-low attachment surface plates with serum-free medium. RNA-sequencing (RNA-seq) was applied in spheroid culture cells and adherent culture cells.
Project description:In vitro proliferation and/or invasion tests (such as MTT assay, Boyden Chamber Assay, Scratch assay) using established and well-characterized cell lines can provide easy and reproducible tools to investigate anti-tumour or anti-invasion agents ex vivo. However, in vivo cells in a tissue context interact with neighbouring cells and extracellular matrix components. In 2-dimensional monolayer cultures cells tend to loose tissue-specific properties. 2-D culture models therefore seem unsuitable to mimic the behaviour of cells in a natural, 3-dimensional microenvironment, especially for testing of growth or invasion inhibiting substances. Spheroids on the other hand resemble in many aspects solid, non-vascularized tumour tissue. Within this study the larynx carcinoma cell line HLaC78 was used as a model for comparing overall gene expression in 2D monolayer culture with 72 hrs old HLaC78 multicellular spheroids, providing evidence for extensive restructuring of gene expression in the 3-dimensional context.
Project description:Mesenchymal stem cells (MSC) are heterogeneous in morphology and transcriptome, resulting in varying therapeutic outcomes. In this study, we found that 3D spheroid culture of heterogeneous MSC, which have undergone conventional 2D monolayer culture for 5-6 passages, synchronized the cells into a uniform cell population with dramatically reduced cell size, and considerably increased levels of immunosuppressive genes and growth factors. Sc-RNA-Seq analysis of the cells revealed that 3D MSC consisted of 2 major cell subpopulations and both expressed high levels of immunosuppressive factors, compared to 6 subpopulations in 2D MSC. In addition, 3D MSC showed a greater suppressive effect on T cells. Moreover, intravenous infusion of a large dose of 3D MSC prior to Imiquimod (IMQ) treatment significantly improved psoriatic lesion. Thus, our results indicate that 3D spheroid culture reprograms heterogeneous mesenchymal stem cells into a uniform immunosuppressive phenotype and promises a novel therapeutic potential for inflammatory diseases.