Dynamic study of In vitro murine follicle maturation in 3% alginate gels
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Beatriz Penalver Bernabe
PROVIDER: E-GEOD-55969 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:A microarray study to compare gene expression during ovarian follicle development to better understand transcriptional changes during follicular growth. Multi-layered secondary follicles (150-180 M-BM-5m in diameter) were mechanically isolated from ovaries of 16-day old CD-1 mice and individually encapsulated in 0.5% alginate. Follicles were culture for up to 8 days. Day 0 samples correspond to follicles that were isolated and immediately frozen. Samples were collected after 0, 2, 4, 5, 6 and 8 days in culture in triplicate and day 0 was collected 5 times.
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Project description:Multilayer murine follicles were grown in 3% alginate and follicles were isolated for transcriptomics analysis at day 0, 2, 4,5,6,and 8 after encapsulation. This data complements to the GSE42795, where follicles were grown in vitro in 0.5% alginate gels
2014-06-01 | GSE55969 | GEO
Project description:We have studied the expression profile of 3D cultured human chondrocytes that were stimulated with supernatant of synovial fibroblasts derived from a RA patient (RASF=HSE cell line) and from a normal donor (NDSF=K4IM cell line), respectively. For this purpose, passage 2 human chondrocytes were cultured for 14 days in alginate beads and subsequently stimulated for 48 hours with supernatant of RASF and NDSF. Baseline expression was determined of unstimulated chondrocytes. Differential genome-wide microarray analysis of RASF and NDSF stimulated chondrocytes disclosed a distinct expression profile related to cartilage destruction involving marker genes of inflammation (COX-2), NF-kappa B signaling pathway (TLR2), cytokines/chemokines and receptors (CXCL1-3, CCL20, CXCL8, CXCR4, IL-6, IL-1beta), matrix degradation (MMP-10, MMP-12) and suppressed matrix synthesis (COMP). Thus, transcriptome profiling of RASF and NDSF stimulated chondrocytes revealed a disturbed catabolic-anabolic homeostasis of chondrocyte function. This study provides a comprehensive insight into the molecular regulatory processes induced in human chondrocytes during RA-related cartilage destruction. Experiment Overall Design: To reveal the RA-related chondrocyte gene expression profile, genome-wide microarray analysis of RASF stimulated human chondrocytes compared to NDSF stimulation was performed. Unstimulated chondrocytes were analyzed for baseline gene expression. For microarray analysis of RASF and NDSF stimulated and unstimulated chondrocytes, respectively, two RNA pools were analyzed, each pool consisting of equal amounts of RNA from three different donors.
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Project description:In this project hiPSC-derived β-like cells were studied in two different 3D enviromnets; size-adjusted cell aggregates (using Aggrewell™ plates) and inside alginate capsules. The hiPSC derived from a patient with maturity-onset diabetes of the young 1 (MODY1), which is a monogenic diabetes form, caused by a mutation in the HNF4A gene. We also generated and included a CRISPR/cas9 corrected isogenic control line. Human islets were also included as positive control. In the data analysis we compared to proteome of the cells in 3D environments compared to the frat 2D enviromnet. We found HNF4A mutation specific effects with both distinct 3D environments, where the cells are challenged differentially in term of oxygen, nutrient supply and cell-to-cell contact. We identified HNF4A mutation-specific phenotypes, including a unique proteome signature suggesting metabolic changes in the alginate bead environment, and mutation-specific cellular phenotypes and a unique proteome signature in the aggregation environment.
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Project description:Rheumatoid arthritis (RA) leads to progressive destruction of articular structures. Despite recent progress in controlling inflammation and pain, little cartilage repair has yet been observed. This in vitro study aims to determine the role of chondrocytes in RA-related cartilage destruction and antirheumatic drug-related regenerative processes. Human chondrocytes were three-dimensionally cultured in alginate beads. To determine the RA-induced gene expression pattern, human chondrocytes were stimulated with supernatant of RA synovial fibroblasts (RASF) and normal donor synovial fibroblasts (NDSF), respectively. To examine antirheumatic drug response signatures, human chondrocytes were stimulated with supernatant of RASF that have been treated with disease-modifying antirheumatic drugs (DMARD; azathioprine, sodium aurothiomalate, chloroquine phosphate, methotrexate), non-steroidal anti-inflammatory drugs (NSAID; piroxicam, diclofenac) or steroidal anti-inflammatory drugs (SAID; methylprednisolone, prednisolone). Genome-wide expression profiling with oligonucleotide microarrays was used to determine differentially expressed genes. Real-time RT-PCR and ELISA were performed for validation of microarray data. Following antirheumatic treatment, microarray analysis disclosed a reverted expression of 94 RA-induced chondrocyte genes involved in inflammation/NF-κB signalling, cytokine/chemokine activity, immune response, proliferation/differentiation and matrix remodelling. Hierarchical clustering analysis showed that treatment of RASF with the DMARD azathioprine, gold sodium thiomalate and methotrexate resulted in chondrocyte gene expression signatures that were closely related to the âhealthyâ pattern. Treatment with the SAID methylprednisolone and prednisolone strongly reverted the RA-related chondrocyte gene expression, in particular the expression of genes involved in inflammation/NF-κB and cytokine/chemokine activity. The NSAID piroxicam and diclofenac and the DMARD chloroquine phosphate had only moderate to marginal effects. Pathway analysis determined major mechanisms of drug action, for example pathways of cytokine-cytokine receptor interaction, TGF-β/TLR/Jak-STAT signalling and ECM-receptor interaction were targeted. This in vitro study provides a comprehensive molecular insight into the antirheumatic drug response signatures in human chondrocytes, thereby revealing potential molecular targets, pathways and mechanisms of drug action involved in chondrocyte regeneration. Thus, the present study may contribute to the development of novel therapeutic chondro-protective compounds and strategies. Experiment Overall Design: Drug-related suppression of gene expression in activated chondrocytes was determined by genome-wide microarray analysis. Chondrocytes were stimulated with supernatant of RASF and NDSF. Effect of treatment with DMARDs, NSAIDs and glucocorticoids was tested by treating RASF prior to collection of supernatant. Two RNA pools were analyzed for each group (RASF-stimulated NDSF stimulated and RASF-treated), each pool consisting of equal amounts of RNA from three different donors.
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Project description:The biocompatibility of immunoisolation devices is a major challenge for their use in cellular therapies. Utilisation of alginate for the encapsulation of pancreatic islets as a potential cellular therapy for type 1 diabetes has shown to be limited in terms of long-term graft survival, due to pericapsular fibrotic overgrowth. Fibrotic overgrowth is a complex process that involves several factors, including protein adsorption. This work investigates the protein adsorption profiles of plasma incubated alginate beads for the identification of key proteins that may play a role in the biocompatibility of this biomaterial.
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Project description:Investigation of whole genome gene expression level changes in Sphingomonas. sp A1 AlgO-deficient mutant grown on alginate compared with that on yeast extract AlgO is a possble transcriptional factor described in J. Bacteriol. (2000) 182(14):3998-4004 by Momma K, Okamoto M, Mishima Y, Mori S, Hashimoto W, and Murata K. A two chip study using total RNA recovered from two cultures of Sphingomonas. sp A1 AlgO-deficient mutant grown in 0.5% alginate medium and 0.5% yeast extract medium. Each chip measures the expression level of genes from Sphingomonas. sp A1.
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