Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab / Generación de osteoclastos a partir de células RAW 264.7 y PBMC. El montaje en nuestro laboratorio

Introduction and objectives: osteoclasts are terminally differentiated giant multinucleated cells derived from the fusionof mononuclear progenitors of the monocyte/macrophage hematopoietic lineage. The objective of our group was to achie-ve 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 differentiationwas assessed by TRAP staining. Osteoclast activity was detected by the resorption pits in Corning® Osteo Assay SurfacePlates.Results: the optimal cell density for RAW 264.7 cell differentiation was 25,000 cells/cm2 with 30 ng/mL of RANKL for6 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.(AU)