Recommended procedure
Reconstitute the bone slices with media before initiating the pit assay. Mature or premature osteoclast
either generated from peripheral blood monocytes (PBMC) or co-cultures can be plated on the bone slices.
Dependent on the maturation state of the osteoclasts and the factors present, resorption pits will appear
after 3-14 days. The number of osteoclasts present can be visualized by its TRAcP activity, while the
formation of resorption pits and trenches can be visualized by toluidine blue staining (see Figure 2).
Figure 1:
Bovine cortical bone slides without human osteoclast.
Figure 2:
Bovine cortical bone slides with human osteoclast for 3 days.
Osteoclastic bone resorption result in the formation of resorption pits which can be used to monitor
the
resorption of the cultured osteoclasts.