Fig. 2: Differentiation potential of sorted trilineage monocyte progenitor (TMP) into macrophages, osteoclasts, and dendritic cells (DCs) under appropriate culture conditions.

A The population of TMPs, defined as CD15⁻CD45⁺CD3⁻CD19⁻CD56⁻CD11b⁺CD14⁺ cells, was labeled and sorted from peripheral blood mononuclear cells (PBMCs) of control subjects. LY; CD3/CD19/CD56. B The re-analysis of sorted TMPs revealed sorting purity of target population above 99.5%. C Experimental design of trilineage differentiation. TMPs were seeded into plates with immobilized Notch ligands (JAG1 or DLL1). Cells were matured in the appropriate culture conditions, and LPS was added (last 24 h) to mimic inflammatory conditions. Functional maturation was assessed in a lineage-specific manner: macrophages were tested by the addition of bacterial particles to assess phagocytosis; osteoclasts were subjected to cytochemical staining for the activity of tartrate-resistant acid phosphatase; DCs were treated with peptide mix and co-cultured with autologous T lymphocytes to evaluate DC capacity to stimulate their proliferation. Created with BioRender.com D Representative brightfield images show typical morphology of differentiated macrophages, osteoclasts, and DCs derived from common TMP (top to bottom; magnification ×100).