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A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

JoVE Journal
Medicina

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JoVE Journal Medicina

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

DOI:

10.3791/64016-v

•

07:03 min

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June 16, 2022

•

Wanjing Cen, Siegmar Reinert, Meltem Avci-Adali, Dorothea Alexander, Felix Umrath³

¹Department of Oral and Maxillofacial Surgery,University Hospital Tübingen, ²Department of Thoracic and Cardiovascular Surgery,University Hospital Tübingen, ³Department of Orthopaedic Surgery,University Hospital Tübingen

Capítulos

00:04Introduction
00:51Calcification of 96-Well Cell Culture Plates
01:54Isolation of Peripheral Blood Mononuclear Cells (PBMCs)
02:49Expansion of Osteoclasts Progenitors and Induction of Osteoclastogenesis
04:19Fluorescence Staining of Osteoclasts and Calcium Phosphate Coating
05:17Results: Analysis of Mature Osteoclast Cultured on Calcium Phosphate Coated Wells
06:38Conclusion

Summary

Tadução automática

Here, we present a simple and effective assay procedure for resorption pit assays using calcium phosphate coated cell culture plates.

Tags

Keywords: Osteoclastic Resorption Calcium Phosphate Coating PBMC Osteoclast Differentiation Bone Marrow Derived Monocytes Raw 264.7 Cells Density Gradient Centrifugation Macrophage Colony Stimulating Factor

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