Project description:The extracellular matrix (ECM)-regulated phenotypic plasticity is crucial for metastatic progression of triple negative breast cancer (TNBC). To investigate the effect of matrix cues on phenotypic plasticity, we embedded triple negative DU4475 cells from suspension culture into mechanically tunable fibrin gels or basement membrane extract (Matrigel). Cells in suspension culture and matrix cultures showed divergent gene expression profiles, with specific gene expression signatures depending on the biochemical composition and stiffness of the matrix.
Project description:Kidney organoids have potential uses in disease modelling, drug screening and regenerative medicine. However, novel cost-effective techniques are needed to enable scale-up production of kidney cell types in vitro. We describe here a modified suspension culture method for the generation of kidney micro-organoids from human pluripotent stem cells. Each micro-organoid contains 6-10 nephrons surrounded by endothelial and stromal populations. Single cell transcriptional profiling confirmed the presence and transcriptional equivalence of all anticipated renal cell types consistent with a previous organoid culture method. Ligand-receptor mapping identified TGFβ signalling between stromal and epithelial cell types as likely to result in fibrotic changes observed with long-term culture. The addition of an ALK4 inhibitor suppressed stromal expansion, maintaining epithelial morphology and improving maturation. This suspension culture micro-organoid methodology resulted in a 3-4-fold increase in final cell yield compared to static culture, thereby representing an economical approach to the production of kidney cells for various biological applications.
Project description:Full title: Three-dimensional culture of AIDS-NHL cells influences gene expression related to B-cell development, proliferation and survival The AIDS-NHL-derived cell line, UMCL01-101, representing diffuse large B-cell lymphoma of immunoblastic morphology (AIDS-IBL), was grown in conventional, static suspension culture or three-dimensionally (3D) in the Rotating Wall Vessel (RWV) bioreactor. The objective was to assess the impact on gene expression of growth as a three-dimensional tissue assembly. Global gene expression analysis was performed on UMCL01-101 cells grown under either condition using Affymetrix microarray. UMCL01-101 cells were cultured in the Rotating Wall Vessel bioreactor to form 3D assemblies, or in conventional suspension culture, for 15 days. RNA was prepared from triplicate samples under each growth condition and submitted for microarray analysis.
Project description:Stage 3 cells from a suspension environment and adherent culture were compared and evaluated through RNA sequencing. RNA sequencing data in heatmap and volcano plot evaluation of core endocrine genes showed that expression levels in adherent cells were lower than those expressed in 3D environment. Several genes that are essential for insulin transcription such as NEUROD1, MAFA, PPARGC1A and NKX2.2 along with INS were expressed with higher transcript levels in bioreactor samples. It was observed that genes usually expressed in earlier stages such as FOXA2 and HNF1b had higher expression levels in adherent culture indicating that 2D environment differentiation might be lagging.
Project description:In this project, trophoblast cell analogue human chorionic carcinoma cell line was cultivated in both conventional monolayer culture (2D) and as spheroids in suspension cultures (3D) and how the cell growth environment affects the physical and biochemical properties of EVs produced by them was studied. Importantly, proteomic cargo profile of EVs derived from 2D and 3D cultures was compared. There was substantial differences in the proteomic cargo profile and potency of EVs derived from two culture systems. Findings of this study will help in selecting an EV production platform, especially for assessing the functional and therapeutic potential of EVs.
Project description:Prospective, open labelled, multicenter trial to evaluate the feasibility of ex vivo culture 3D (chemogram obtaining) on biopsies in order to estimate the predictive value of this technique for treatment response in patients treated by two different chemotherapies (FOLFOX or FOLFIRI) for colorectal cancer.