Isolation and Culture of Human Osteoblasts

Gartland, Alison; Rumney, Robin M. H.; Dillon, J.P.; Gallagher, James A.

doi:10.1007/978-1-61779-367-7_22

Cited by 42 publications

(32 citation statements)

References 18 publications

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“…Bone : Pieces of cancellous bone were harvested with a curette and subjected to enzymatic digestion with 0.1% collagenase II and 0.2% trypsin (BioConcept, Allschwil, Switzerland) at 37 °C on an orbital shaker for 1.5 h. The digested bony pieces were placed in a Petri dish containing bone culture medium consisting of DMEM/F-12 medium supplemented with 10% FBS, 1% penicillin/streptomycin, 0.2% Amphotericin B (Thermo Fisher Scientific) and 0.5 μg/mL ascorbic acid (Sigma Aldrich, Buchs, Switzerland) [28]. Isolated cells were transferred to culture flasks containing the bone culture medium as described above.…”

Section: Methodsmentioning

confidence: 99%

Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing

Kuemmerle

Theiss

Okoniewski

et al. 2016

Genes

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Although several tendon-selective genes exist, they are also expressed in other musculoskeletal tissues. As cell and tissue engineering is reliant on specific molecular markers to discriminate between cell types, tendon-specific genes need to be identified. In order to accomplish this, we have used RNA sequencing (RNA-seq) to compare gene expression between tendon, bone, cartilage and ligament from horses. We identified several tendon-selective gene markers, and established eyes absent homolog 2 (EYA2) and a G-protein regulated inducer of neurite outgrowth 3 (GPRIN3) as specific tendon markers using RT-qPCR. Equine tendon cells cultured as three-dimensional spheroids expressed significantly greater levels of EYA2 than GPRIN3, and stained positively for EYA2 using immunohistochemistry. EYA2 was also found in fibroblast-like cells within the tendon tissue matrix and in cells localized to the vascular endothelium. In summary, we have identified EYA2 and GPRIN3 as specific molecular markers of equine tendon as compared to bone, cartilage and ligament, and provide evidence for the use of EYA2 as an additional marker for tendon cells in vitro.

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Section: Methodsmentioning

confidence: 99%

Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing

Kuemmerle

Theiss

Okoniewski

et al. 2016

Genes

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“…Media was changed every 2 to 3 days and differentiation was assessed at 7 and 14 days after treatment began by examining alkaline phosphatase (Alp) and at 14 and 21 days using alizarin red staining to detect mineralization, as described previously. (48) Osteosarcoma SAOS-2 cells (ATCC, Teddington, UK) were cultured as described previously, (49) treated as above for 3 days, and cultured in 1% FCS to measure Alp. Each experiment was repeated at least three times (n ¼ 3) and representative data are presented as a fold change.…”

Section: Osteoblast Isolation and Differentiationmentioning

confidence: 99%

Preventing and Repairing Myeloma Bone Disease by Combining Conventional Antiresorptive Treatment With a Bone Anabolic Agent in Murine Models

Paton-Hough

Tazzyman

Evans

et al. 2018

Journal of Bone and Mineral Research

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Multiple myeloma is a plasma cell malignancy, which develops in the bone marrow and frequently leads to severe bone destruction. Current antiresorptive therapies to treat the bone disease do little to repair damaged bone; therefore, new treatment strategies incorporating bone anabolic therapies are urgently required. We hypothesized that combination therapy using the standard of care antiresorptive zoledronic acid (Zol) with a bone anabolic (anti‐TGFβ/1D11) would be more effective at treating myeloma‐induced bone disease than Zol therapy alone. JJN3 myeloma‐bearing mice ( n = 8/group) treated with combined Zol and 1D11 resulted in a 48% increase ( p ≤ 0.001) in trabecular bone volume (BV/TV) compared with Zol alone and a 65% increase ( p ≤ 0.0001) compared with 1D11 alone. Our most significant finding was the substantial repair of U266‐induced osteolytic bone lesions with combination therapy ( n = 8/group), which resulted in a significant reduction in lesion area compared with vehicle ( p ≤ 0.01) or Zol alone ( p ≤ 0.01). These results demonstrate that combined antiresorptive and bone anabolic therapy is significantly more effective at preventing myeloma‐induced bone disease than Zol alone. Furthermore, we demonstrate that combined therapy is able to repair established myelomatous bone lesions. This is a highly translational strategy that could significantly improve bone outcomes and quality of life for patients with myeloma. © 2018 The Authors. Journal of Bone and Mineral Research Published by Wiley Periodicals Inc.

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“…As the single cellular constituent of adult articular cartilage, chondrocytes are responsible for maintaining the cartilage matrix at a low turnover state of equilibrium. In mature cartilage, chondrocytes synthesize small proteoglycans such as biglycan and decorin and other specific and non‐specific matrix proteins very slowly [Gartland et al, 2005].…”

mentioning

confidence: 99%

Proteomic and redox‐proteomic evaluation of homogentisic acid and ascorbic acid effects on human articular chondrocytes

Braconi

Laschi

Taylor

et al. 2010

J of Cellular Biochemistry

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Alkaptonuria (AKU) is a rare genetic disease associated with the accumulation of homogentisic acid (HGA) and its oxidized/polymerized products in connective tissues up to the deposition of melanin-like pigments (ochronosis). Since little is known on the effects of HGA and its metabolites on articular cells, we carried out a proteomic and redox-proteomic analysis to investigate how HGA and ascorbic acid (ASC) affect the human chondrocytic protein repertoire. We settled up an in vitro model using a human chondrocytic cell line to evaluate the effects of 0.33 mM HGA, alone or combined with ASC. We found that HGA and ASC significantly affect the levels of proteins with specific functions in protein folding, cell organization and, notably, stress response and cell defense. Increased protein carbonyls levels were found either in HGA or ASC treated cells, and evidences produced in this paper support the hypothesis that HGA-induced stress might be mediated by protein oxidation. Our finding can lay the basis towards the settling up of more sophisticated models to study AKU and ochronosis.

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Isolation and Culture of Human Osteoblasts

Cited by 42 publications

References 18 publications

Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing

Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing

Preventing and Repairing Myeloma Bone Disease by Combining Conventional Antiresorptive Treatment With a Bone Anabolic Agent in Murine Models

Proteomic and redox‐proteomic evaluation of homogentisic acid and ascorbic acid effects on human articular chondrocytes

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