Optimization of the isolation and differentiation protocol of human bone marrow derived osteoblasts and osteoclasts

last updated: 2020-01-10
ProjectFROnTHERA :: publications list
TitleOptimization of the isolation and differentiation protocol of human bone marrow derived osteoblasts and osteoclasts
Publication TypeComunications - Poster
Year of Publication2018
AuthorsBastos A. R., Maia F. R., Oliveira J. M., Reis R. L., and Correlo V. M.

Bone tissue has a remarkable capacity to self-regenerate in case of damage promoted due to several diseases or trauma. Nevertheless, bone tissue loses this capacity when the damage exceeds a critical size [1]. Autologous bone grafts are still the gold standard treatment. Nevertheless, limited supply and donor site complications are the biggest limitations. Tissue engineering (TE) strategies have been emerged as promising alternatives to these restrictions. Nevertheless, in most studies, osteoblasts (OB) are the main type of bone cell studied, disregarding the importance of osteoclasts (OC). But, in fact, bone is a complex tissue that depends not only on OB for tissue formation, but also OC for tissue resorption. Thus, TE strategies should consider both cell populations, OB and OC, to obtain a fully functional bone tissue.

This can be due to the fact that only few researchers have successfully generated large numbers of OC from human precursor cells, in contrary to the generation of OB, as described in the literature [2].

In this context, we propose an experimental setup for the isolation and differentiation of, not only, OB, but also, OC from human bone marrow (hBM). For that, an optimized protocol for the collection of hBM, described by Cody, J. J. et al. will be follow [3]. Further, hBM will be submitted to different methods to isolate OB and OC precursor’s cells: (i) Cells’ adhesion; (ii) Cell sorting; and (iii) Cell sorting through magnetic microbeads. Finally, the optimal conditions to achieve a successful differentiation of OB and OC will be pursued. OB precursor’s cells will be culture under osteogenic inducers (ascorbic acid, β-glycerophosphate and dexamethasone), while OC precursors will be cultured under osteoclastogenic inducers (M-CSF and RANKL). To access the proper differentiation, different markers will be used. Overall, we expect that this study leads to important findings, improving the current protocols tested/used by the scientific community.

Conference NameFinal Conference Chem2Nature
Date Published2018-10-24
Conference Location Centro Cultural Vila Flor, em Guimarães.
Keywordsbone remodeling, human bone marrow
Peer reviewedno

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