Flow Perfusion Co-culture of Human Mesenchymal Stem Cells and Endothelial Cells on Biodegradable Polymer Scaffolds

In this study, we investigated the effect of flow perfusion culture on the mineralization of co-cultures of human umbilical vein endothelial cells (HUVECs) and human mesenchymal stem cells (hMSCs). Osteogenically precultured hMSCs were seeded onto electrospun scaffolds in monoculture or a 1:1 ratio with HUVECs, cultured for 7 or 14 days in osteogenic medium under static or flow perfusion conditions, and the resulting constructs were analyzed for cellularity, alkaline phosphatase (ALP) activity and calcium content. In flow perfusion, constructs with monocultures of hMSCs demonstrated higher cellularity and calcium content, but lower ALP activity compared to corresponding static controls. ALP activity was enhanced in co-cultures under flow perfusion conditions, compared to hMSCs alone; however unlike the static controls, the calcium content of the co-cultures in flow perfusion was not different from the corresponding hMSC monocultures. The data suggest that co-cultures of hMSCs and HUVECs did not contribute to enhanced mineralization compared to hMSCs alone under the flow perfusion conditions investigated in this study. However, flow perfusion culture resulted in an enhanced spatial distribution of cells and matrix compared to static cultures, which were limited to a thin surface layer

Enhanced osteogenesis in co-cultures with human mesenchymal stem cells and endothelial cells on polymeric microfiber scaffolds

In this work, human mesenchymal stem cells (hMSCs) and their osteogenically precultured derivatives were directly co-cultured with human umbilical vein endothelial cells (HUVECs) on electrospun 3D poly(-caprolactone) microfiber scaffolds in order to evaluate the co-culture’s effect on the generation of osteogenic constructs. Specifically, cells were cultured on scaffolds for up to three weeks, and the cellularity, alkaline phosphatase activity (ALP), and bone-like matrix formation were assessed. Constructs with co-cultures and monocultures had almost identical cellularity after the first week, however lower cellularity was observed in co-cultures compared to monocultures during the subsequent two weeks of culture. Scaffolds with co-cultures showed significantly higher ALP activity, glycosaminoglycan and collagen production, as well as greater calcium deposition over the course of study compared to monocultures of hMSCs. Furthermore, the osteogenic outcome was equally robust in co-cultures containing osteogenically precultured and non-precultured hMSCs. The results demonstrate that the combination of MSC and HUVEC populations within a porous scaffold material under osteogenic culture conditions is an effective strategy to promote osteogenesis