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Osteoblasts

Osteoblasts are the bone-forming cells responsible for the synthesis and mineralization of the extracellular matrix in bone tissue.
They originate from mesenchymal stem cells and play a crucial role in bone development, growth, and remodeling.
Osteoblasts secrete collagen, proteoglycans, and other organic components that form the osteoid, which subsequently undergoes mineralization to create the hard, calcified matrix of bone.
These cells also regulate the activity of osteoclasts, the bone-resorbing cells, through the production of signaling molecules.
Proper osteoblast function is essential for maintaining bone homeostasis and preventing skeletal disorders such as osteoporosis.
Reasearch on osteoblasts is crucial for understanding bone physiology and developing therapies for bone-related diseases.

Most cited protocols related to «Osteoblasts»

Osteocyte-like phenotype culture

Tissue culture media were purchased from GIBCO BRL, fetal bovine serum (FBS) was from BioWhittaker. Rat tail collagen type 1, 99% pure, was purchased from Becton Dickinson Laboratories. All other reagents were purchased from Sigma Chemical Co. unless otherwise stated. Cells were expanded in permissive conditions (33°C in αMEM with 10% FBS, 100 units/ml penicillin, 50 µg/ml streptomycin, and 50 U/ml IFN-γ) on rat tail type I collagen-coated plates or gels or bovine type I collagen sponges. To induce osteogenesis, cells were plated at 80,000 cells/cm² in osteogenic conditions (37°C with 50 µg/ml ascorbic acid and 4 mM β-glycerophosphate in the absence of IFN-γ). Collagen-coated surfaces were used because they were found to be effective at maintaining an osteocyte-like phenotype (10 (link)).
MLO-A5 cells, used as controls, are an established model of late osteoblasts with the ability to rapidly synthesize mineralized extracellular matrix (1 (link)). MLO-A5 cells are highly responsive to mechanical loading in 3D culture (15 (link)). MLO-Y4 cells, also used as controls, are an established model of osteocytes.

Woo S.M., Rosser J., Dusevich V., Kalajzic I, & Bonewald L.F. (2011). Cell Line IDG-SW3 Replicates Osteoblast-to-Late-Osteocyte Differentiation in vitro and Accelerates Bone Formation in vivo. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 26(11), 2634-2646.

Publication 2011

Ascorbic Acid beta-glycerol phosphate Bos taurus Cells Collagen Collagen Type I Culture Media Extracellular Matrix Fetal Bovine Serum Gels Interferon Type II Osteoblasts Osteocytes Osteogenesis Penicillins Phenotype Porifera Streptomycin Tail Tissues

ATAC-seq and Capture C Analysis

The ATAC-seq and capture C methods have been previously published [15 (link)]. Briefly, we first identified all proxy SNPs in LD (r² = 0.5) with the sentinel GWAS SNPs using raggr (www.raggr.usc.edu) with the following parameters: populations: CEU + FIN+GBR + IBS + TSI; min MAF = 0.001; min r² = 0.5; max r² = 1.0; max distance = 500 kb; max Mendelian errors = 1; HWP cutoff = 0; min genotype % = 75; genotype database = 1000 Genomes Phase 3; genome build hg19. We then assessed which of these proxy SNPs and which of the gene promoters baited in our capture C library resided in an open chromatin region in hMSC-derived osteoblasts, by intersecting their genomic positions with those of the ATAC-seq peaks (using the BEDTools function intersectBed with 1 bp overlap). Next, we extracted the chromatin loops linking open proxy SNPs and open gene promoters in the hMSC-derived osteoblast capture C dataset (bait-to-bait interactions were excluded). The results were visualized using the UCSC Genome Browser.

Cousminer D.L., Wagley Y., Pippin J.A., Elhakeem A., Way G.P., Pahl M.C., McCormack S.E., Chesi A., Mitchell J.A., Kindler J.M., Baird D., Hartley A., Howe L., Kalkwarf H.J., Lappe J.M., Lu S., Leonard M.E., Johnson M.E., Hakonarson H., Gilsanz V., Shepherd J.A., Oberfield S.E., Greene C.S., Kelly A., Lawlor D.A., Voight B.F., Wells A.D., Zemel B.S., Hankenson K.D, & Grant S.F. (2021). Genome-wide association study implicates novel loci and reveals candidate effector genes for longitudinal pediatric bone accrual. Genome Biology, 22, 1.

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Publication 2021

ATAC-Seq Chromatin DNA Library Genome Genome-Wide Association Study Genotype Osteoblasts Population Group Promoter, Genetic Single Nucleotide Polymorphism

Zebrafish Caudal Fin Regeneration

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Publication 2012

Adult Amputation Animals, Transgenic CCL4 protein, human Cells Codon Cre recombinase Embryo Enzymes Escherichia coli Estrogen Receptors Ethanol Fishes Genes Hemizygote Homo sapiens Hybrids Nitroreductases Open Reading Frames Oryzias latipes Osteoblasts Osteocalcin Plasmids Strains Tamoxifen tricaine Zebrafish

Genetic Manipulation of Notch Signaling in Bone

For wild-type bone analysis, C57BL/6J male mice were used unless stated otherwise. All EC-specific gene targeting experiments were performed with Cdh5(PAC)-CreERT2 transgenic mice24 (link). For Rbpj deletion in the postnatal endothelium, mice carrying loxP-flanked Rbpj (Rbpj^lox/lox) alleles25 (link) and Cdh5(PAC)-CreERT2 transgenics were interbred. To induce Cre activity and gene deletion, offspring was injected with 500µg tamoxifen (Sigma, T5648) intraperitoneally every day from P10 to P14. The resulting Rbpj^iΔEC (CreERT2^T/+Rbpj^lox/lox) mutants and Cre- littermate controls were sacrificed at P28, and femurs and tibiae were collected for analysis. Identical breeding and tamoxifen administration strategies were used to generate EC-specific mutants with Fbxw7^lox/lox (Ref.10 (link)), Dll1^lox/lox (Ref. 13 (link)), Dll4^lox/lox (Ref. 14 (link)), or Jag1^lox/lox (Ref. 15 (link)) mice and inducible osteoblast-specific Rbpj knockouts (Rbpj^iOB). The latter were generated with Tg(Col1a1-creERT2)6.1.ICS transgenic mice (Institut Clinique de la Souris, France).
For Notch gain-of-function experiments, Gt(ROSA)26Sor^{tm1(Notch1)Dam/J} mice carrying a Cre-inducible transgene for Notch1 intracellular domain overexpression26 (link) and Cdh5(PAC)-CreERT2 transgenics were interbred. Tamoxifen administration (see above for injection schedule) was used to generate CreERT2-positive (NICD^iOE-EC) mutants overexpressing NICD in ECs and controls. For experiments with EC-specific Dll4^iΔEC/NICD^iOE-EC double mutants, interbreeding of Dll4^lox/lox conditional mice14 (link) with Gt(ROSA)26Sor^{tm1(Notch1)Dam/J} and Cdh5(PAC)-CreERT2 transgenics generated triple heterozygote males, which were bred to Dll4^lox/+Gt(ROSA)26Sor^{tm1(Notch1)Dam/J} double heterozygous females. This produced CreERT2-negative controls together with Dll4^lox/lox NICD^+/+CreERT2^T/+ (Dll4^iΔEC) and Dll4^+/+ NICD^+/OECreERT2^T/+ (NICD^iOE-EC) single mutants, and Dll4^iΔEC/NICD^iOE-EC double mutants, which received tamoxifen and were analysed as described above. For mutant analysis, both male and female mice were used.
For Noggin administration experiments, mice were injected intraperitoneally once daily with 500µg/kg recombinant Noggin (R&D Systems) from P15 to P27, after completion of tamoxifen injections (P10-P14) and before analysis at P28.
All animal experiments were performed in compliance with the relevant laws and institutional guidelines and were approved by local animal ethics committees.

Ramasamy S.K., Kusumbe A.P., Wang L, & Adams R.H. (2014). Endothelial Notch activity promotes angiogenesis and osteogenesis in bone. Nature, 507(7492), 376-380.

Publication 2014

Animal Ethics Committees Animals, Transgenic Bones CDH5 protein, human Deletion Mutation Endothelium Females Femur Gene Deletion Heterozygote Males Mice, Inbred C57BL Mice, Laboratory Mice, Transgenic noggin protein Osteoblasts Protoplasm RBPJ protein, human Rosa Tamoxifen Tibia Transgenes

Comprehensive ChIP-Seq and DNase-Seq Profiling

We chose to run our segmentation (see below) on a pre-selected set of ChIP-Seq assays (CTCF, H3K4me1, H3K4me2, H3K4me3, H3K9ac, H3K27ac, H3K27me3, H3K36me3, H4K20me1) along with DNaseI hypersensitivity and a control ChIP-Seq experiment. We therefore downloaded from ENCODE 2 and Epigenomics Roadmap all the raw read datasets produced by ChIP-Seq and DNaseI hypersensitivity experiments on the 18 cell types that had all of the above required assays: A549, DND-41, GM12878, H1-hESC, HeLa-S3, HepG2, HMEC, HSMM, HSMMtube, HUVEC, IMR90, K562, Monocytes-CD14+, NH-A, NHDF-AD, NHEK, NHLF, Osteoblast. Including replicates and control samples, this amounted to 740 datasets, all referenced in the Ensembl homo_sapiens_funcgen_76_38 MySQL database.

Zerbino D.R., Wilder S.P., Johnson N., Juettemann T, & Flicek P.R. (2015). The Ensembl Regulatory Build. Genome Biology, 16(1), 56.

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Publication 2015

Biological Assay Cells Chromatin Immunoprecipitation Sequencing CTCF protein, human HeLa Cells histone H3 trimethyl Lys4 Homo sapiens Human Embryonic Stem Cells Hypersensitivity Immunoprecipitation, Chromatin Monocytes Osteoblasts

Most recents protocols related to «Osteoblasts»

Immunosuppressive Tumor-Derived MSCs

Not available on PMC !

Example 7

Tumor-Derived MSC-Like Lymphoma Stromal Cells are Immunosuppressive

Since the tumor cells in lymphoma are not adherent, it is possible to isolate tumor stromal cells from lymphomas developed in p53+/− mice. It was observed that these cells can be passaged in vitro and can be differentiated into adipocytes and osteoblast-like cells. Interestingly, like bone marrow derived MSCs, these tumor stromal cells are also immunosuppressive and can effectively inhibit the proliferation of ant-CD3-activated splenocytes. This immunosuppressive effect was also dependent on IFNγ+TNF α and NO, since anti-IFNγ IFNγ and iNOS inhibitors could reverse the immunosuppressive effect.

US11872250B2. Methods for inducing an immune response by administering activated mesenchymal stem cells (2024-01-16). Rutgers, The State University of New Jersey [US]. Inventors: Yufang Shi [US], Guangwen Ren [US], Liying Zhang [US].

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Patent 2024

Adipocytes Bone Marrow Cardiac Arrest Cells Immunosuppressive Agents inhibitors Interferon Type II Lymphoma Mesenchyma Mus Neoplasms NOS2A protein, human Osteoblasts Response, Immune Stem, Plant Stromal Cells Tumor-Derived Activated Cells Tumor Necrosis Factor-alpha

Chitosan Nanofibers for Tissue Engineering

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Example 5

To further explore the potential of the electrospun chitosan nanofibers in tissue engineering applications, osteoblast proliferation on the membrane of electrospun chitosan nanofibers was examined by Celltiter Glo Assay Kit. As shown in FIG. 12, there was no statistical difference in the day 5 growth of cells on chitosan compared with acylated (acetyl- and butyryl-) and deacylated electrospun chitosan nanofiber membranes, suggesting the electrospun chitosan nanofibers membranes were non-toxic.

The cell morphology on the materials was visualized with fluorescence microscope after osteoblast cells were cultured on the top of the materials for 5 days. The cells grown on the electrospun chitosan nanofibers showed characteristic shapes associated with osteoblast cells, such as elongated/stretched shape, suggesting the material did not interfere with the growth of the osteoblasts.

These surprising results suggested that the problems with dissolution and swelling observed with electrospun chitosan fiber membranes can be solved by the reversible acylation method. The mechanisms behind the process were elucidated based on the data obtained from the FTIR, XPS and SEM analyses. The acylation method could potentially be used to synthesize other modified chitoan nanofibrous material containing acyl moieties as well.

US11878088B2. Chitosan nanofiber compositions, compositions comprising modified chitosan, and methods of use (2024-01-23). The University of Memphis Research Foundation [US]. Inventors: Joel D. Bumgardner [US], Hengjie Su [US], Tomoko Fujiwara [US], Daniel G. Abebe [US], Kwei-Yu Liu [US].

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Patent 2024

Acylation Biological Assay Cell Proliferation Cells Chitosan Fibrosis Microscopy, Fluorescence Osteoblasts Somatostatin-Secreting Cells Spectroscopy, Fourier Transform Infrared Tissue, Membrane

Quantitative Analysis of Early and Late Mineralization

The early mineralization was determined based on the instructions of BCIP/NBT alkaline phosphatase chromogenic kit (Beyotime, Shanghai, China). Cells were fixed in 4% paraformaldehyde (Beyotime) for 10 min, washed three times with PBS, and then incubated with BCIP/NBT staining solution for 30 min at 37 °C in the dark. After that, the treated cells were washed twice with distilled water and photographed. Then, ALP concentration was quantitatively analyzed. Late mineralization was measured according to the instructions of the Osteoblast Mineralized Nodule Staining Kit (Beyotime). The cells were fixed with 4% paraformaldehyde for 10 min, washed with PBS for another three times, and stained with alizarin red for 10 min. Subsequently, the treated cells were washed with distilled water to be observed and photographed microscopically, and the calcium nodules level was then quantitatively analyzed.

Jiang J., Zhang N., Song H., Yang Y., Li J, & Hu X. (2023). Oridonin alleviates the inhibitory effect of lipopolysaccharide on the proliferation and osteogenic potential of periodontal ligament stem cells by inhibiting endoplasmic reticulum stress and NF-κB/NLRP3 inflammasome signaling. BMC Oral Health, 23, 137.

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Publication 2023

azo rubin S Calcium Cells Osteoblasts paraform Physiologic Calcification

Visualizing Osteoblast-Exosome Interactions

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Publication 2023

Actins Cell Nucleus Coculture Techniques DAPI Exosomes Fluorescein-5-isothiocyanate Fluorescence Fluorescent Dyes Macrophage Microscopy, Fluorescence Osteoblasts Osteoclasts

Isolation and culture of murine primary osteoblasts

Murine primary osteoblasts were obtained as described in previous research [24 (link), 25 (link)]. Briefly, the calvarial bone was carefully dissected from one-day-old neonatal Wistar rats and washed with PBS solutions containing 100 U/ml penicillin and 100 µg/ml streptomycin thrice. The semi-transparent calvarial bone was incubated with 0.25% trypsin at 37 °C for thirty minutes, then sliced and incubated with 0.1% collagenase I at 37 °C overnight. The isolated osteoblasts were collected from the final digests and resuspended in the complete medium consisting of α-MEM, 10% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin. Cells were cultured in the humidified atmosphere of 5% CO₂.

Xiao X., Zou S, & Chen J. (2023). Cyclic tensile force modifies calvarial osteoblast function via the interplay between ERK1/2 and STAT3. BMC Molecular and Cell Biology, 24, 9.

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Publication 2023

Atmosphere Bones Calvaria Cells Collagenase, Clostridium histolyticum collagenase 1 Infant, Newborn Mus Osteoblasts Penicillins Rats, Wistar Streptomycin Trypsin

Top products related to «Osteoblasts»

Fetal bovine serum (fbs) by Thermo Fisher Scientific

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific

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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.

β glycerophosphate by Merck Group

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β-glycerophosphate is a chemical compound that serves as a buffering agent and source of phosphate for cell culture media. It helps maintain a stable pH environment for cell growth and proliferation.

α mem by Thermo Fisher Scientific

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α-MEM is a cell culture medium formulated for the growth and maintenance of mammalian cells. It provides a balanced salt solution, amino acids, vitamins, and other nutrients required for cell proliferation.

Dexamethasone (dex) by Merck Group

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Dexamethasone is a synthetic glucocorticoid medication used in a variety of medical applications. It is primarily used as an anti-inflammatory and immunosuppressant agent.

Penicillin streptomycin by Thermo Fisher Scientific

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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.

Ascorbic acid by Merck Group

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Ascorbic acid is a chemical compound commonly known as Vitamin C. It is a water-soluble vitamin that plays a role in various physiological processes. As a laboratory product, ascorbic acid is used as a reducing agent, antioxidant, and pH regulator in various applications.

Penicillin by Thermo Fisher Scientific

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Penicillin is a type of antibiotic used in laboratory settings. It is a broad-spectrum antimicrobial agent effective against a variety of bacteria. Penicillin functions by disrupting the bacterial cell wall, leading to cell death.

Hfob1 by American Type Culture Collection

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The HFOB1.19 is a laboratory equipment piece designed for cell culture applications. It provides a controlled environment for the growth and maintenance of cells. The HFOB1.19 is capable of regulating temperature, humidity, and gas composition to support optimal cell growth conditions.

Streptomycin by Thermo Fisher Scientific

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Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It functions as a protein synthesis inhibitor, targeting the 30S subunit of bacterial ribosomes, which plays a crucial role in the translation of genetic information into proteins. Streptomycin is commonly used in microbiological research and applications that require selective inhibition of bacterial growth.

More about "Osteoblasts"

Osteoblasts are the primary bone-forming cells, responsible for the synthesis and mineralization of the extracellular matrix in bone tissue.
These specialized cells originate from mesenchymal stem cells and play a crucial role in bone development, growth, and remodeling processes.
Osteoblasts secrete key components like collagen, proteoglycans, and other organic materials that form the osteoid, which then undergoes mineralization to create the hard, calcified bone matrix.
These cells also regulate the activity of osteoclasts, the bone-resorbing cells, through the production of signaling molecules.
Proper osteoblast function is essential for maintaining bone homeostasis and preventing skeletal disorders such as osteoporosis.
Researchers studying osteoblasts often utilize various cell culture media and supplements, including fetal bovine serum (FBS), Dulbecco's Modified Eagle Medium (DMEM), β-glycerophosphate, α-Minimal Essential Medium (α-MEM), dexamethasone, penicillin/streptomycin, and ascorbic acid, to support osteoblast growth and differentiation in vitro.
Cell lines like HFOB1.19 are commonly used as model systems to investigate osteoblast biology and physiology.
Understanding the complex mechanisms governing osteoblast function is crucial for developing effective therapies for bone-related diseases and promoting overall skeletal health.