Effects of the conditioned medium on TMD and BMD spheroids. Of note, MC = MC3T3 osteoblast-like cells, A5 = MLO-A5 osteocyte-like cells, and Y4 = MLO-Y4 osteocyte-like cells. The asterisk indicates statistical significance at p < 0.05. (A) Primary cell spheroids formed in media conditioned with MLO-A5 and MLO-Y4 spheroids after 24 h. (B) Roughness and area of the primary cell spheroids. (C & D) TMD and BMD spheroids in the conditioned media isolated from MC3T3, MLO-A5, and MLO-Y4 cultures. (E & F) Relative size change of TMD and BMD spheroids in the conditioned media.