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380 results for “Chondrocytes”
Analysis of hMSC-derived chondrocytes in response to stimulation with SARS-CoV-2 spike proteins
<p>Previous studies have detected the presence of SARS-CoV-2 viral proteins in bronchial cartilage chondrocytes. We hypothesised that the leakage of viral proteins to local joint tissue was due to virus-induced endothelial dysfunction. Studies have also shown upregulation of endothelin-1 (ET-1), the most potent vasoconstrictor, in COVID patients. We are investigating the direct effect of SARS-CoV-2 spike protein (SP) and the host response to chondrocytes.</p> <p>Human mesenchymal stem cells (hMSCs)-differentiated chondrocytes were treated with either full-length SARS-CoV-2 spike protein (SP) only or a combination of spike protein, neutralising antibody to S1 and endothelin-1 (SAE) to mimic viral insult and host response respectively. RNA sequencing was performed to compared the change in transcriptome in control, SP, and SAE. All samples were processed in the same batch. Default quality control parameters were used.</p>
Biomarker Signatures of Quality for Engineering Nasal Chondrocyte-Derived Cartilage
<p>Data underlying the figures in the publication “Biomarker Signatures of Quality for Engineering Nasal Chondrocyte-Derived Cartilage”, published in <em>Front Bioeng Biotechnol., </em><strong>2020</strong>, 8: 283. (DOI: 10.3389/fbioe.2020.00283).</p> <p>Table of contents:</p> <p><strong>1. biochem_summary</strong>; Biochemistry data and MBS of pellets made from NC and PC mixtures in the titration experiment.</p> <p><strong>2. dirty-clean-biochem</strong>; Biochemistry data and MBS of pellets made from passage 2 cells from dirty/clean biopsies.</p> <p><strong>3. identity_all</strong>; All passage 1 and passage 2 cells of type pure NC and pure PC from the titration and dirty/clean datasets. All PCR data included.</p> <p><strong>4. identity_titration</strong>; Passage 1 and 2 and pellets made from pure NC and PC populations in the titration experiment. All PCR data included. Average (avg) refers to the average of the biologicsl replicates (which are already the average of the experimental replicates of Data 5).</p> <p><strong>5. identity_titration_avg</strong>; Passage 1 and 2 and pellets made from pure NC and PC populations in the titration experiment. All PCR data included. All experimental replicate information is included (which are the average of the technical replicates).</p> <p><strong>6. PCR_cells_summary</strong>; For the purity and potency assays, mixtures of increasing NC to PC cell ratios were created. The PCR data from those mixtures are here for passage 2 and for the pure populations as passage 1.</p> <p><strong>7. PCR_pellet_summary</strong>; For the purity and potency assays, mixtures of increasing NC to PC cell ratios were created. PCR data from engineered pellets are here included.</p> <p><strong>8. PCRdirty-clean</strong>; PCR data of cells obtained from dirty/clean biopsies at passage 0,1, and 2.</p> <p><strong>9. PCRsummary_all</strong>; Combined Data 5, 7, and 8.</p> <p><strong>10. Proliferation_TOT</strong>; Proliferation rates of cells from clean and dirty biopsies. </p>
Confocal microscopy images (CZI files) of human chondrocytes of different resolutions and magnifications with stained nuclei and primary cilia
<p>This dataset is an addition of https://doi.org/10.5281/zenodo.7994589. Here, we have focused on the influence of the magnifications of microscope objectives and image resolution on the results of automated cilia length measurements.</p> <p>1. Methods</p> <p>1.1 Cell culture</p> <p>Human non-degenerative chondrocytes from a 30-year-old male donor (NHAC-kn, CC-2550; LONZA, Walkersville Inc., Walkersville, MD, USA) were used. These chondrocytes were seeded in passage four with a density of 28000 cells/cm<sup>2</sup> on collagen-coated glass coverslips (GG-15-Collagen; Neuvitro Corporation, Camas, WA, USA). The cells were cultivated in 12-well plates (Thermo Fisher Scientific Inc., Waltham, MA, USA) under hypoxic conditions at 37°C, 5% CO<sub>2</sub> and 5% O<sub>2</sub> with different media compositions for three days.<br> The basal medium consisted of Dulbecco’s Modified Eagle Medium (DMEM) (Gibco™) including high glucose (GlutaMAX™), sodium pyruvate supplements (Thermo Fisher Scientific Inc., Waltham, MA, USA), as well as 1% penicillin/streptomycin (Pen/Strep; Thermo Fisher Scientific Inc.), 1% Amphotericin B (Biochrom GmbH, Berlin, Germany), and 50 µg mL<sup>−1</sup> ascorbic acid (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany). To this basal medium, the following supplements were added: ITS with Dexa + IGF-1 + TGF-β1: 1% Insulin-Transferrin-Selenium (ITS+™), 100 nM dexamethasone, 50 ng mL<sup>−1</sup> insulin-like growth factor (IGF)-1 (R&D Systems, Minneapolis, MN, USA) and 50 ng mL<sup>−1</sup> transforming growth factor (TGF)-β1 (Peprotec, Hamburg, Germany).</p> <p>1.2 Immunocytochemistry</p> <p>After three days of cultivation in the different media compositions, the chondrocytes were washed once with phosphate-buffered saline (PBS; Biochrom GmbH, Berlin, Germany) and fixed for 10 min at room temperature (RT) with 4% paraformaldehyde (ROTI ® Histofix, Carl Roth GmbH + Co. KG, Karlsruhe, Germany). After fixation, cells were washed again and permeabilized with 0.2% Triton-X100 (Merck, Darmstadt, Germany) for 10 min. For blocking the unspecific binding sites, cell-seeded coverslips were incubated with bovine serum albumin (BSA; Sigma-Aldrich) with a concentration of 5% in PBS for one hour at RT after another washing step with PBS. To stain the primary cilium, cells were incubated with anti-acetylated α-tubulin (6-11B-1) (RRID: AB 628409) labeled with Alexa Fluor 647 (sc-23950 AF647, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:200 in PBS at 4°C overnight. Additionally, the Actin cytoskeleton was stained with Acti-stain 488 Fluorescent Phalloidin (Cytoskeleton, Inc., Denver, CO, USA) diluted 10 in PBS for 30 min at RT. Afterward, cells were washed three times with PBS, and the coverslips were fixed with Fluoroshield™ (Sigma-Aldrich) containing 4’,6-Diamidino-2-phenylindole (DAPI).</p> <p>1.3 Image acquisition</p> <p>Three-dimensional fluorescence images of stained cells were acquired with a ZEISS ELYRA LSM 780 confocal laser scanning microscope (CLSM) (Carl Zeiss AG, Oberkochen, Germany). To find optimal microscopy parameters for automated detection and length measurement of primary cilia, images were recorded using a Plan-Apochromat 63×/1.40 Oil DIC M27 objective (Carl Zeiss AG, Oberkochen, Germany) or an α Plan-Apochromat 100 × /1.46 Oil DIC M27 Elyra objective (Carl Zeiss AG, Oberkochen, Germany) as well as the following resolutions: 1024 × 1024, 2048 × 2048 and 4096 × 4096 pixels resulting in different voxel sizes (see metadata of files).</p>
Confocal microscopy images (CZI files) of human chondrocytes in different cell culture media with stained nuclei and primary cilia
<p>1. Methods</p> <p>1.1 Cell culture</p> <p>For investigating the influence of the cell culture medium composition on the lengths of primary cilia, human non-degenerative chondrocytes from a 30-year-old male donor (NHAC-kn, CC-2550; LONZA, Walkersville Inc., Walkersville, MD, USA) were used. These chondrocytes were seeded in passage four with a density of 28000 cells/cm<sup>2</sup> on collagen-coated glass coverslips (GG-15-Collagen; Neuvitro Corporation, Camas, WA, USA). The cells were cultivated in 12-well plates (Thermo Fisher Scientific Inc., Waltham, MA, USA) under hypoxic conditions at 37°C, 5% CO<sub>2</sub> and 5% O<sub>2</sub> with different media compositions for three days.<br> The basal medium consisted of Dulbecco’s Modified Eagle Medium (DMEM) (Gibco™) including high glucose (GlutaMAX™), sodium pyruvate supplements (Thermo Fisher Scientific Inc., Waltham, MA, USA), as well as 1% penicillin/streptomycin (Pen/Strep; Thermo Fisher Scientific Inc.), 1% Amphotericin B (Biochrom GmbH, Berlin, Germany), and 50 µg mL<sup>−1</sup> ascorbic acid (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany). To this basal medium, different supplements were added, creating four groups:<br> 1) ITS: 1% Insulin-Transferrin-Selenium (ITS+™ Premix, BD Biosciences, Franklin Lakes, NJ, USA),<br> 2) ITS with Dexa: 1% Insulin-Transferrin-Selenium (ITS+™) and 100 nM dexamethasone (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany),<br> 3) ITS with Dexa + IGF-1 + TGF-β1: 1% Insulin-Transferrin-Selenium (ITS+™), 100 nM dexamethasone, 50 ng mL<sup>−1</sup> insulin-like growth factor (IGF)-1 (R&D Systems, Minneapolis, MN, USA) and 50 ng mL<sup>−1</sup> transforming growth factor (TGF)-β1 (Peprotec, Hamburg, Germany),<br> 4) FBS: 10% fetal bovine serum (FBS, Pan Biotech, Aidenbach, Germany).</p> <p>1.2 Immunocytochemistry</p> <p>After three days of cultivation in the different media compositions, the chondrocytes were washed once with phosphate-buffered saline (PBS; Biochrom GmbH, Berlin, Germany) and fixed for 10 min at room temperature (RT) with 4% paraformaldehyde (ROTI ® Histofix, Carl Roth GmbH + Co. KG, Karlsruhe, Germany). After fixation, cells were washed again and permeabilized with 0.2% Triton-X100 (Merck, Darmstadt, Germany) for 10 min. For blocking the unspecific binding sites, cell-seeded coverslips were incubated with bovine serum albumin (BSA; Sigma-Aldrich) with a concentration of 5% in PBS for one hour at RT after another washing step with PBS. To stain the primary cilium, cells were incubated with anti-acetylated α-tubulin (6-11B-1) (RRID: AB 628409) labeled with Alexa Fluor 647 (sc-23950 AF647, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:200 in PBS at 4°C overnight. Additionally, the Actin cytoskeleton was stained with Acti-stain 488 Fluorescent Phalloidin (Cytoskeleton, Inc., Denver, CO, USA) diluted 10 in PBS for 30 min at RT. Afterward, cells were washed three times with PBS, and the coverslips were fixed with Fluoroshield™ (Sigma-Aldrich) containing 4’,6-Diamidino-2-phenylindole (DAPI).</p> <p>1.3 Image acquisition</p> <p>Three-dimensional fluorescence images of stained cells were acquired with a ZEISS ELYRA LSM 780 confocal laser scanning microscope (CLSM) (Carl Zeiss AG, Oberkochen, Germany). Images were recorded using a Plan-Apochromat 63×/1.40 Oil DIC M27 objective (Carl Zeiss AG, Oberkochen, Germany). The distance of two layers was 0.2814 µm and the resolution 1024 × 1024 pixels (scan magnification: 0.6, pixel length: 0.2196 µm).</p>
The Protective Function of Directed Asymmetry in the Pericellular Matrix Enveloping Chondrocytes (Supporting Data)
<p>This dataset contains the images necessary to reproduce the study "The Protective Function of Directed Asymmetry in the Pericellular Matrix Enveloping Chondrocytes (DOI: 10.1007/s10439-021-02900-1)"</p>
An in vivo stable isotope labeling method to investigate individual matrix protein synthesis, ribosomal biogenesis, and chondrocyte proliferation in murine articular cartilage
<p>These experiments have used a stable-isotope method using <em>in vivo</em> deuterium oxide labeling and mass spectrometry to measure protein concentration, protein half-life, cell proliferation, and ribosomal biogenesis in a single sample of murine articular cartilage. We hypothesized that a 60-day labeling period would capture age-related declines in cartilage matrix protein content, protein synthesis rates, and chondrocyte proliferation. Knee cartilage was isolated from 25- and 90-week-old female C57BL/6J mice treated with deuterium oxide for 15, 30, 45 and 60 days. We measured protein abundance and half-lives using high resolution accurate mass spectrometry (HRAM) and d2ome data processing software. </p>
Data for: NFATc1 marks articular cartilage progenitors and negatively determines articular chondrocyte differentiation
<p>The origin and differentiation mechanism of articular chondrocytes remain poorly understood. Broadly, the difference in developmental mechanisms of articular and growth-plate cartilage is still less elucidated. Here, we identified that the nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) is a crucial regulator of articular, but not growth-plate, chondrocyte differentiation during development. At the early stage of mouse knee development (embryonic day 13.5), NFATc1-expressing cells were mainly located in the flanking region of the joint interzone. With development, NFATc1-expressing cells generated almost all articular chondrocytes, but not chondrocytes in limb growth-plate primordium. NFATc1-expressing cells displayed prominent capacities for colony formation and multipotent differentiation. Transcriptome analyses revealed a set of characteristic genes in NFATc1-enriched articular cartilage progenitors. Strikingly, the expression of NFATc1 was diminished with articular chondrocyte differentiation and suppressing NFATc1 expression in articular cartilage progenitors was sufficient to induce spontaneous chondrogenesis while overexpressing NFATc1 suppresses chondrogenesis. Mechanistically, NFATc1 negatively regulated the transcriptional activity of the <em>Col2a1</em> gene. Thus, our results reveal that NFATc1 characterizes articular, but not growth-plate, cartilage progenitors during development and negatively determines articular chondrocyte differentiation at least partly through regulating COL2A1 gene transcription.</p>
Data for: NFATc1 marks articular cartilage progenitors and negatively determines articular chondrocyte differentiation
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Lin28a induces SOX9 and chondrocyte reprogramming via HMGA2 and blunts cartilage loss in mice
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Connexin 43 Modulation in Human Chondrocytes, Osteoblasts and Cartilage Explants: Implications for Inflammatory Joint Disorders
<p>Dataset of the scientific publication "Connexin 43 Modulation in Human Chondrocytes, Osteoblasts and Cartilage Explants: Implications for Inflammatory Joint Disorders"</p>
Autologous Chondrocyte Transplantation For Articular Cartilage Regeneration
ClinicalTrials.gov study NCT01503970. IPD Sharing: Not stated. Countries: 1. Publications: 3.
Study on Safety, Tolerability and Prelimenary Efficacy of LNA043 in Patients Undergoing Autologous Chondrocyte Implantation
ClinicalTrials.gov study NCT03334812. IPD Sharing: YES. Countries: 1. Publications: 0.
Comparison of Autologous Chondrocyte Implantation Versus Mosaicoplasty: a Randomized Trial
ClinicalTrials.gov study NCT00560664. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Safety and Efficacy Study of Cells Sheet-Autologous Chondrocyte Implantation to Treat Articular Cartilage Defects
ClinicalTrials.gov study NCT01694823. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Arthroscopic Autologous Chondrocyte Implantation Versus Microfractures
ClinicalTrials.gov study NCT01947374. IPD Sharing: Not stated. Countries: 1. Publications: 4.
Autologous Mesenchymal Stem Cells vs. Chondrocytes for the Repair of Chondral Knee Defects
ClinicalTrials.gov study NCT01399749. IPD Sharing: Not stated. Countries: 1. Publications: 3.
Autologous Chondrocyte Implantation in the Patellofemoral Joint
ClinicalTrials.gov study NCT00212849. IPD Sharing: Not stated. Countries: 1. Publications: 1.
RNA sequencing data of gene expression of human chondrocytes cultured at 33℃ or 37 ℃
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Long-term Outcome Evaluation of Patients Undergoing Autologous Chondrocyte Transplantation Delivered on Biomaterial
ClinicalTrials.gov study NCT06238947. IPD Sharing: Not stated. Countries: 0. Publications: 10.
Effects of CHONDRON (Autologous Chondrocytes) With Ankle Cartilage Defect
ClinicalTrials.gov study NCT01050816. IPD Sharing: Not stated. Countries: 1. Publications: 0.
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