Figure 1.

Experimental setup illustrating the temporal approach used to induce the osteogenic differentiation of hASCs and consequent establishment of a direct co‐culture model and correspondent single cultures. To assess the influence of different concentrations of osteogenic supplementation on culture medium, both pre‐OBs and hTDCs were initially seeded separately in single culture systems. For this purpose, cellular responses were studied to simultaneously evaluate the effects on the osteogenic commitment of pre‐OBs and the tenogenic phenotype of hTDCs. Subsequently, co‐culture systems were established using pre‐OBs and hTDCs under direct contact conditions to further investigate the crosstalk occurring between these 2 cell types under selected culture medium conditions. In this case, single culture systems were also performed in parallel as controls