Table 3.

Main strategies of coculturing osteogenic and vascular precursors for vascularized bone engineering

Medium composition	Cell ratio (O:V)	Seeding methodology
Osteogenic/vascular [88, 298]	1:1 [299]	Simultaneous seeding [229, 300, 301]
Expansion/vascular [227]	1:4 [301]	Pros: technically simpler; even mix distribution within the construct; cell–cell crosstalk during differentiation	Cons: risk of suboptimal individual cell type viability and differentiation
Osteogenic [228]	2:1 [88]	Sequential seeding [89]
Expansion [302]	4:1 [301]	Pros: optimal differentiation of the first seeded cells; cell–cell communication	Cons: uneven distribution of the two types of cells within the construct
Vascular [300]	5:1 [302]	Independent differentiation [88]
Osteogenic with vascular growth factors [229]	8:1 [303]	Pros: optimal differentiation of each cell type in their respective medium	Cons: lack of cell–cell communication
	3:2 [300]

O osteogenic, V vascular, Expansion expansion medium for osteogenic precursors