Gene expression during primary osteoblast differentiation
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Thomas Streichert
PROVIDER: E-GEOD-71565 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Primary murine osteoblasts were isolated form the calvariae of newborn mice. 10 days after the addition of ascorbic acid (50 µg/ml) and ß-glycerophosphate (10 mM), cells were serum-starved over night and then incubated for 6 hours with condtioned medium of MDA-PCa2b cells or conditioned medium of PC-3 cells Keywords: expression profiling by array
2010-07-01 | GSE17348 | GEO
Project description:Primary murine osteoblasts were isolated form the calvariae of newborn mice. 10 days after the addition of ascorbic acid (50 µg/ml) and Ã-glycerophosphate (10 mM), cells were serum-starved over night and then incubated for 6 hours with condtioned medium of MDA-PCa2b cells or conditioned medium of PC-3 cells Keywords: expression profiling by array The aim of the experiment was to identify transcriptional changes in osteoblasts caused by factors derived from two different prostate cancer cell lines
2010-07-30 | E-GEOD-17348 | biostudies-arrayexpress
Project description:Mucolipidosis type II (MLII) is a severe inherited multisystemic disorder caused by mutations in the GNPTAB gene. Skeletal abnormalities are a predominant feature of MLII. Here we investigate the gene expression in a knock-in mouse model for mucolipidosis type II, generated by the insertion of a cytosine in the murine Gnptab gene (c.3082insC) that is homologous to a homozygous mutation in an MLII patient. Since osteoblasts are critically involved in regulating bone development and remodeling, a genome-wide expression analysis was performed with RNA isolated from primary cultures of osteoblasts originating from MLII knock-in mice (KI) compared to RNA from wild-type (WT) osteoblasts to identify dysregulated genes involved in pathogenic mechanisms. Primary osteoblasts were isolated from calvaria of 5-day-old wild-type (WT) and MLII knock-in littermates (KI). RNA was extracted at day 10 of differentiation induced by ascorbic acid and beta-glycerophosphate and hybridization on Affymetrix microarrays. We used preparations of RNA from two individual primary cultures of osteoblasts for every genotype (WT_OB_I, WT_OB_II, KI_OB_I, KI_OB_II) and compared WT vs KI samples.
2014-01-29 | E-GEOD-43854 | biostudies-arrayexpress
Project description:MC3T3 osteoblasts were grown in culture medium in the presence of 10 mM beta-glycerophosphate and and 100 micro g per mL of ascorbic acid and 25% LLC1 conditioned medium or vehicle for 7 days. Cells were lysed an RNA was isolated.
2023-04-15 | GSE152513 | GEO
Project description:Bone mineralization is a carefully orchestrated process, regulated by a number of promoters and inhibitors that function to ensure effective hydroxyapatite formation. Here we sought to identify new regulators of this process through a time series microarray analysis of mineralising primary osteoblast cultures over a 27 day culture period.
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Project description:The cytosolic protein Sharpin is as a component of the linear ubiquitin chain assembly complex (LUBAC), which regulates NF-κB signaling in response to specific ligands. Its inactivating mutation in Cpdm (chronic proliferative dermatitis mutation) mice causes multi-organ inflammation, yet this phenotype is not transferable into wildtype mice by hematopoietic stem cell transfer. Recent evidence demonstrated that Cpdm mice additionally display low bone mass, but the cellular and molecular causes of this phenotype remained to be established. Here we have applied non-decalcified histology together with cellular and dynamic histomorphometry to perform a thorough skeletal phenotyping of Cpdm mice. We show that Cpdm mice display trabecular and cortical osteopenia, solely explained by impaired bone formation, whereas osteoclastogenesis is unaffected. We additionally found that Cpdm mice display a severe disturbance of articular cartilage integrity in the absence of joint inflammation, supporting the concept that Sharpin-deficiency affects mesenchymal cell differentiation. Consistently, Cpdm mesenchymal cells displayed reduced osteogenic capacitiy ex vivo, yet this defect was not associated with impaired NF-κB signaling. A molecular comparison of wildtype and Cpdm bone marrow cell populations further revealed that Cpdm mesenchymal cells produce higher levels of Cxcl5 and lower levels of IL1ra. Collectively, our data demonstrate that skeletal defects of Cpdm mice are not caused by chronic inflammation, but that Sharpin is as a critical regulator of mesenchymal cell differentiation and gene expression. They additionally provide an alternative molecular explanation for the inflammatory phenotype of Cpdm mice and the absence of disease transfer by hematopoetic stem cell transplantation. Unsorted bone marrow cells from wildtype and Cpdm mice were cultured for 10 days in the presence of ascorbic acid and ß-glycerophosphate to induce osteogenic differentiation
2015-09-07 | E-GEOD-61299 | biostudies-arrayexpress
Project description:We recently identified Bicc1 as a regulator of osteoblast differentiation and bone mass. Bicc1 encodes an RNA-binding protein. Here, we have used siRNA to decrease Bicc1 expression in primary calvarial osteoblasts. Illlumina microarrays were then used to profile global gene expression changes. The experiment consisted of 16 total samples in 4 treatment groups (N=4 replicates per treatment). The treatment groups were osteoblasts treated with 1). control scrambled siRNA, 2) Bicc1_1 siRNA, 3) Bicc1_2 siRNA and 3) Bicc1_3 siRNA. Cells were isolated, transfected and 48 hours later RNA was isolated and hybridized to arrays.
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Project description:The osteogenic differentiation process consists in a well-coordinated multi-step sequence of events, in which each stage is characterized by the expression of distinct protein markers, as well as by individual morphological features. In order to precisely identify the step at which cells are permissive to MET-driven transformation, we used expression profiling. We compared the transcriptional profiles of MET-HOB and MSC cells where osteoblast differentiation was induced. Using microarrays, we evaluated the expression profiles of MET-HOB and MSC cells where osteoblast differentiation was induced, by culturing cells in the presence of ascorbic acid, α-glycerophosphate and dexamethasone for 7, 14 and 21 days
2013-02-28 | E-GEOD-28205 | biostudies-arrayexpress
Project description:Rat osteoblasts ROB17/2.8 cells were cultured in the presence of high (1mg/ml) or low (0.01mg/ml) concentration of polyphosphates. Poly(P) induced gene expression in rat osteoblast ROS17/2.8 cells was measured after the treatment with high (1mg/ml) or low (0.01mg/ml) for 24 h.
2016-03-15 | E-GEOD-79176 | biostudies-arrayexpress
Project description:In the present study, the Agilent-016251 Sparus aurata oligo microarray platform (GEO accession: GPL6467) was used to compare expression profiles of mineralization-induced VSa16 cell cultures against untreated ones. ECM mineralization was induced for 4 weeks by supplementing medium with 50 µg/ml of L-ascorbic acid, 10 mM β glycerophosphate and 4 mM CaCl2. For each group, total RNA was extracted from three (3) independent biological replicates, each consisting of pools of cells. Data analysis demonstrated that expression profiles were strongly affected by ECM mineralization with hundreds of genes differentially expressed with relevant fold-change. In this study, we analyzed six (6) cell samples, three (3) collected from untreated VSa16 cell cultures and three (3) collected from mineralization-induced VSa16 cell cultures. Gene expression profiling was performed using Agilent-016251 Sparus aurata oligo microarray platform (GPL6467) (6 arrays, no replicate) based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. Feature Extraction (FE) 9.5 used XDR ID to link the pairs of scans together automatically when extracting data. The signal that was left after all the FE processing steps was ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal.
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