Tunneling nanotubes mediate intercellular communication between endothelial progenitor cells and osteoclast precursors

Tunneling nanotube (TNT)-mediated cell communication play pivotal roles in a series of physiological and pathological processes in multicellular organism. This study was designed to investigate the existence of TNTs between EPCs and osteoclast precursors and evaluate their effects on the differentiation of osteoclast precursors. For these purposes, EPCs and osteoclast precursors (RAW264.7 cells) were stained with different fluorescent dyes before direct co-culture; then, the co-cultured cells were sorted by fluorescence activated cell sorter (FACS), and the differentiation of co-cultured RAW264.7 cells was evaluated. The results showed that the differentiation potential of RAW264.7 cells was significantly inhibited after their co-culture with EPCs. Additionally, the expression of macrophage migration inhibitory factor (MIF) was up-regulated in RAW264.7 cells after co-culture. Moreover, the MIF inhibitor ISO-1 could rescue the formation of TRAP-positive multinuclear osteoclasts and the expression of osteoclastogenesis-associated genes in the co-cultured RAW264.7 cells. The present study demonstrates that EPCs can affect the differentiation of osteoclast precursors through the TNT-like structures formed across these two types of cells and might inform new therapeutic strategies for osteolytic diseases.