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Differentiation of Functional Osteoclasts from Human Peripheral Blood CD14+ Monocytes
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An Optimized Method to Generate Human Active Osteoclasts From Peripheral Blood Monocytes.

Dina Abdallah1, Marie-Laure Jourdain1, Julien Braux1

  • 1EA 4691 "Biomatériaux et Inflammation en site osseux" SFR CAP-Santé (FED 4231), Université Reims Champagne-Ardenne, Reims, France.

Frontiers in Immunology
|April 20, 2018
PubMed
Summary

This study optimized osteoclast (OC) generation from human PBMCs. A 14-day culture with M-CSF, RANKL, and charcoal-stripped serum efficiently produces active human OCs for research.

Keywords:
CD14+ monocytesdentin matrix resorptionhuman peripheral blood mononuclear cellsosteoclastsvitamin D3

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Area of Science:

  • Cell Biology
  • Immunology
  • Bone Biology

Background:

  • Osteoclasts (OCs) are crucial for bone remodeling and skeletal development.
  • OCs are implicated in bone disorder pathogenesis.
  • Generating mature human OCs from peripheral blood mononuclear cells (PBMCs) presents technical challenges.

Purpose of the Study:

  • To develop an optimized procedure for generating active, mature human OCs from fresh PBMCs.
  • To establish a standardized source of human OCs for research.

Main Methods:

  • PBMCs were cultured in various media with M-CSF and RANKL.
  • Charcoal-stripped and non-stripped sera were compared.
  • Effects of vitamin D3 were investigated.
  • OC differentiation, morphology, and resorption activity were assessed over 14-21 days using fluorescent staining and scanning electron microscopy.

Main Results:

  • An optimized 14-day culture protocol was established.
  • The optimal medium included M-CSF, RANKL, and fetal bovine charcoal-stripped serum.
  • Vitamin D3 was found to be unnecessary for efficient OC differentiation.
  • The method allows efficient production of active human OCs from low PBMC seeding densities.

Conclusions:

  • An efficient and standardized method for generating active human osteoclasts from PBMCs has been developed.
  • This optimized protocol facilitates further research into human OC function and bone-related diseases.