TY  - JOUR
TI  - Pit Assay to Measure the Bone Resorptive Activity of Bone Marrow-derived Osteoclasts
AU  - Vesprey, Alexander 
AU  - Yang, Wentian 
VL  - 6
IS  - 12
PY  - 2016
DA  - 2016/06/20
SP  - e1836
C1  - Bio-protocol 2016;6:e1836
DO  - 10.21769/BioProtoc.1836
UR  - https://doi.org/10.21769/BioProtoc.1836
AB  - Although it is possible to use a tartrate-resistant acid phosphatase (TRAP) stain to assist in identifying osteoclasts, a separate method is needed to determine the bone resorption activity of osteoclasts. Since osteoclasts leave “pits” after bone matrix resorption (Charles et al., 2014), it is possible to stain pits as a method of measuring osteoclast bone resorption activity. The pit assay protocol enables researchers to stain bony slices that were co-cultured with osteoclasts with toluidine blue in order to allow the visualization, capture, and analysis of osteoclast resorptive activity based on the number, size and depth of pits (Zhou et al., 2015). The pit assay protocol is separated into three sequential stages: Preparation of bone slices (1); preparation of osteoclast precursors (Ross et al., 2006; Teitelbaum et al., 2000) (2), and bone resorption pit assay (3).
KW  - Osteoclast function
KW  - Bone resorption
KW  - Pit assay
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False