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Revista de Osteoporosis y Metabolismo Mineral
versão On-line ISSN 2173-2345versão impressa ISSN 1889-836X
Resumo
JURADO, Susana et al. Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab. Rev Osteoporos Metab Miner [online]. 2023, vol.15, n.1, pp.6-11. Epub 29-Maio-2023. ISSN 2173-2345. https://dx.doi.org/10.20960/revosteoporosmetabminer.00005.
Introduction and objectives:
osteoclasts are terminally differentiated giant multinucleated cells derived from the fusion of mononuclear progenitors of the monocyte/macrophage hematopoietic lineage. The objective of our group was to achieve the best method for osteoclast differentiation, from both RAW 264.7 cells and peripheral blood monocytes.
Material and methods:
RAW 264.7 cells and human PBMCs were differentiated into osteoclasts. Success in differentiation was assessed by TRAP staining. Osteoclast activity was detected by the resorption pits in Corning® Osteo Assay Surface Plates.
Results:
the optimal cell density for RAW 264.7 cell differentiation was 25,000 cells/cm2 with 30 ng/mL of RANKL for 6 days. Osteoclasts differentiated from PBMCs were observed after 4 weeks with 25 ng/mL M-CSF and 30 ng/mL RANKL. Individual pits or multiple pit clusters were observed on the surface plates.
Conclusions:
we report optimal conditions for the differentiation of osteoclasts from
Palavras-chave : Osteoclasts; Differentiation; Bone resorption.
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