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Abstract

Medicine

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

Published: June 16th, 2022

DOI:

10.3791/64016

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

ERRATUM NOTICE

Important: There has been an erratum issued for this article. Read more …

Abstract

Mature osteoclasts are multinucleated cells that can degrade bone through the secretion of acids and enzymes. They play a crucial role in various diseases (e.g., osteoporosis and bone cancer) and are therefore important objects of research. In vitro, their activity can be analyzed by the formation of resorption pits. In this protocol, we describe a simple pit assay method using calcium phosphate (CaP) coated cell culture plates, which can be easily visualized and quantified. Osteoclast precursors derived from human peripheral blood mononuclear cells (PBMCs) were cultured on the coated plates in the presence of osteoclastogenic stimuli. After 9 days of incubation, osteoclasts were fixed and stained for fluorescence imaging while the CaP coating was counterstained by calcein. To quantify the resorbed area, the CaP coating on plates was stained with 5% AgNO3 and visualized by brightfield imaging. The resorption pit area was quantified using ImageJ.

Erratum

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

This corrects the article 10.3791/64016

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Keywords Osteoclastic Resorption

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