3D Bioprinting of HGFs and HUVECs Co-culture System Promotes Vascularization of HUVECs
Objective:To construct a co-culture system of human gingival fibroblasts(HGFs)and human umbilical vein endothelial cells(HUVECs)to promote vascularization of HUVECs.Methods:HGFs and HUVECs were cul-tured to 3 to 5 passages for experiments.The HUVECs was labeled with a red fluorescent protein by lentiviral transfection.A two-dimensional co-culture system of HGFs and HUVECs was constructed.HGFs and HUVECs were co-cultured at different ratios(4∶1,1∶1,and 1∶4),and the formation of vascular network was observed under fluorescence microscope.5％methacrylylated gelatin(GelMA)30 hydrogel was prepared to construct a three-dimensional co-culture system of HGFs &HUVECs,and the formation of vascular network was observed un-der laser confocal microscope and quantitative analyzed.HUVECs were separated from three-dimensional co-culture system to assess the proliferation,migration,tube formation,and expression levels of relevant angiogenesis genes after single-culture and co-culture for a certain period.Results:In two-dimensional co-culture,vascular network for-mation was best when HGFs and HUVECs were cocultured in a ratio of 1∶1.In three-dimensional co-culture,HGFs can promote HUVECs angiogenesis.The co-culture of HGFs & HUVECs had better effects on prolifera-tion,migration,and tube formation compared to the three-dimensional culture of HUVECs alone,and the expres-sion levels of relevant angiogenesis genes were significantly higher.Conclusion:Co-culture of HGFs and HUVECs can guide and promote HUVECs migration and sprouting.
3D bioprintingvascularizationtissue en-gineeringhuman umbilical vein endothelial cellshuman gingival fibroblasts
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