Osteoclasts are conventionally visualized by histochemical staining followed by manual counting, which hinders kinetic studies and automated quantification. Here we describe two fluorescence-based assays for the real-time analysis of myeloid cell to osteoclast development (FRAMCO) in primary mouse bone marrow cell cultures.

2023年12月7日 · We report a demineralized bone paper (DBP)-based osteoblast culture and osteoclast assay platform that replicates osteoclast fusion, fission, resorption, and apoptosis with high fidelity and...

Fluorescent staining of osteoclasts to assess F-actin ring formation. The effects of the different inhibitor treatments on osteoclast differentiation on bovine bone slices were studied by fluorescent staining of nuclei and F-actin rings.

The following assays are commonly used to characterize osteoclasts. Staining for tartrate-resistant acid phosphatase. Multinucleated osteoclasts are characterized by high expression of the enzyme tartrate-resistant acidic phosphatase as previously described in Coxon et al.,18 and Itzstein and van't Hof.27 This can easily be stained for and ...

2024年7月8日 · Immunofluorescence staining was employed to examine the spatial distribution and quantity of osteoclast cell skeletons, P2X7 protein, and cell nuclei, while pit assay was used to evaluate osteoclast bone resorption ability.

In this method article, we propose a TRAP activity staining technique suitable for long-term preservation and three-dimensional (3D) imaging. Paraffin sections of decalcified mouse bone are prepared, stained with NewFuchsin TRAP, coverslipped in a non-aqueous mounting medium, and examined (Fig. 1). Fig. 1.

5 天之前 · (F, G) TRAP staining and quantification of osteoclasts treated with FSH (0, 10, and 20 ng/ml) (n = 5). Scale bar = 200 μm. (H) F-actin staining. Scale bar = 200 μm. (I) Quantification of nuclei per osteoclast (n = 5). ... This suggests that Mdh2 may be a critical regulatory factor in osteoclast differentiation.

2024年7月1日 · Collectively, our findings revealed a novel stage (OSC) within the osteoclast lineage, potentially representing elusive bone reversal cells due to its increased chemotactic ability towards MSCs and potential contribution to bone remodelling.

2 天之前 · Osteoporosis (OP) is a systemic disease characterized by low bone mass. New progress has been made in the study of OP, such as lipid peroxidation. However, the role of lipid peroxides in osteoclast differentiation is still unclear. Bone marrow macrophages (BMMs) were extracted from C57BL/6J mice and induced to differentiate into osteoclasts, which were observed via TRAP staining, Phalloidin ...

1 天前 · Subsequently, we assessed osteoclast formation through TRAcP staining and Giemsa fluorescence staining. The results showed that in the lncRNA-MIR193BHG knockdown (Si-Lnc) group, the number of osteoclasts was significantly reduced, whereas in the DNMT3A overexpression (Ov-DNMT3A) group, the number of osteoclasts was significantly increased.