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Figure 4. Co-culture of PMs and cells. (A) CLSM images of EO-MT and C-MT were cultured

under different culture conditions for different time (Blue: DAPI). Quantification of adhesion and

proliferation of the cells in (B, C) EO-MTs and (D, E) C-MTs under static and dynamic culture

modes (n=3).

In normal subchondral bone, endothelial cells are essential in intercellular signaling and

angiogenesis, as well as osteogenesis. Hence, the strategy to prepare EO-MTs apparently mimics

the subchondral tissue. The co-culturing of endothelial cells and osteocytes are the most direct

approach to establish the vascular functions of supplying nutrients and metabolite excretion toward

the construction of bone tissue models. Accordingly, MAEC cells were cocultured with MC3T3-

E1 using PMs, mimicking the native vascularized bone tissue. To investigate the distribution of the

cells in EO-MT, the osteoblasts and the endothelial cells were fluorescently labeled by

Qtracker®655 and Qtracker®525, respectively. Although the angiogenic sprouting and vascular

lumen formation are rarely seen, CLSM images indicated that both the cells, MAEC and MC3T3-

E1, were distributed in the microtissues with high cell viabilities (Figure 5A). Together, the

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