ABSTRACT: To study the impact of the organotypic assembly of vascular smooth muscle cells on their transcriptome, we cultured human umbilical artery smooth muscle cells under 2D conditions and as aggregates in hanging drops under 3D conditions. After 48 hours, RNA was isolated from both groups
Project description:To study the impact of TGF signaling on the transcriptome of vascular smooth muscle cells, we cultured human umbilical artery smooth muscle cells as 3D aggregates in hanging drops for 48 hours. One group was treated with TGFβRI/II inhibitor LY2109761 (10 μM, additional 10 μM after 24 h), the other group with solvent (DMSO) as control. RNA was isolated from both groups after 48h
Project description:To understand the the effect of microparticles conjugated with vascular endothelial growth factor (VEGF) on human endothelial cells derived from human umbilical cord blood (UCB) CD34+ hematopoietic stem cells, we have employed microRNA microarray profiling. The microparticles used in this study were purchased from Invitrogen (Dynal magnetic microparticles coated with streptavidin, 4.5 micron in size) and modified in our lab. Cell suspensions were mixed with blank microparticles (without VEGF), soluble VEGF or VEGF-conjugated microparticles and seeded as hanging drops to prepare endothelial cell aggregates. Cell aggregates without any treatments were used as control. After 2 h or 8 h, the cell aggregates were collected and miRNA expression profiles were detected.
Project description:The current objectives are to use of transcriptome sequencing analysis technology comprehensively understand the potential molecular mechanisms between vascular smooth muscle cells grown in 2D environments (2D-VSMCs) and vascular smooth muscle cells grown in 3D PGA environments (3D-VSMCs).These findings help in gaining a better grasp of the situation of how VSMCs survive on PGA scaffolds and have guiding significance for TEBV culture.
Project description:Human bone marrow mesenchymal stromal cells (MSCs) are conventionally cultured as adherent monolayers on tissue culture plastic. MSCs can also be cultured as 3D cell aggregates (spheroids). Optimised 3D conditions (60,000 MSCs cultured as a spheroid for 5 days) inhibited MSC proliferation and induced cell shrinkage in the absence of cell death. Primary human MSCs isolated from 2 donors were cultured under both monolayer (2D MSCs) and optimised 3D (3D MSCs) conditions. High quality RNA was isolated from all samples, and global gene expression analysis was performed in duplicate (using Agilent SurePrint G3 Human Gene Expression 8x60K v2 Microarrays) to identify gene expression changes in 3D compared to 2D MSC cultures.
Project description:The experiment is part of a project to study DNA repair process after ionizing radiation in organotypic 3-dimentional human bronchial epithlial cell culture. Human bronchial epithelial cells were grown in tissue culture flask (2D) or in matrics gel (3D). Three independent cultures were done for each condition.
Project description:To profile changes in gene expression in human endothelial cells in response to VEGF-A165 and phenotypic changes during vascular network formation in vitro 16 samples of human umbilical vein endothelial cells were analysized, four distinct biological samples were used in each condition. The four conditions included: VEGF treatment in Matrigel culture, Mock treatment in Matrigel culture, VEGF treatment in 2D monolayer, and mock treatment in 2D monolayer.
Project description:To evaluate the effects of FEDB on NCCs induced in 2D conditions, we sampled 2D cultured cells and cell aggregates during FEDB treatment and compared gene expression patterns.
Project description:Tridimensional cardiac differentiation from hiPSCs has been largely described in the literature. However, the exact impact that 3D culture has throughout the entire process of cardiac differentiation remains poorly defined. We developed a robust and efficient 3D platform for cardiomyocyte differentiation from hiPSCs, based on the temporal modulation of WNT signalling using small molecules. 3D aggregates of hiPSCs were generated by forced aggregation in microwells and subsequently differentiated. In order to determine the differences in gene expression profile due to 3D culture throughout the different stages of cardiac differentiation, we compared transcriptional changes between cells in 3D aggregates and standard 2D monolayer cardiac differentiation. Analysis of these data suggests a faster commitment of hiPSCs toward the cardiac lineage and also higher degree of cardiomyocyte functional maturation after 20 days of culture in the 3D aggregates when compared with the 2D monolayer.
Project description:Cell samples of undifferentiated human umbilical cord mesenchymal stem cells (1-3) and cells that have been cultured in smooth muscle differentiation medium for 6 hours (4-6) and 24 hours (7-9) were collected and subjected to miRNA array. Exploration of miRNA involved smooth muscle differentiation mechanism would offer potential therapeutic choices for improving performance of vascular grafts engineered with umbilical cord mesenchymal stem cells.