3D生物打印构建HGFs和HUVECs共培养体系促进HUVECs成血管
3D Bioprinting of HGFs and HUVECs Co-culture System Promotes Vascularization of HUVECs
李俊俊 1王雯 2郭慧颖 1袁长永 2夏廷旭 1王鹏来2
作者信息
- 1. 徐州医科大学口腔医学院 江苏徐州 221004
- 2. 徐州医科大学口腔医学院 江苏徐州 221004;徐州医科大学附属口腔医院种植科 江苏徐州 221000
- 折叠
摘要
目的:构建人牙龈成纤维细胞(human gingival fibroblasts,HGFs)和人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)共培养体系,促进HUVECs成血管.方法:取HGFs和HUVECs培养传代至3～5代进行实验.通过慢病毒转染用红色荧光蛋白标记HUVECs.构建HGFs和HUVECs 二维共培养体系,将HGFs与HUVECs以不同比例(4∶1、1∶1、1∶4)共培养,荧光显微镜下观察血管网络形成情况.配制5％甲基丙烯酰化明胶(GelMA)30胶,构建HGFs和HUVECs三维共培养体系,激光共聚焦显微镜下观察血管网络的形成情况.对HUVECs形成的血管网络进行定量分析.将HUVECs从三维共培养系统中分离出来,评估单独培养和共培养一定时间后HUVECs的增殖、迁移和小管形成效果以及相关血管生成基因的表达水平.结果:二维共培养时,当HGFs和HUVECs以比例1∶1共培养时血管网络形成的效果最佳.三维共培养时,HGFs可以促进HUVECs血管生成.HGFs和HUVECs共培养组的增殖、迁移和小管形成效果更好,相关血管生成基因的表达水平远远高于HUVECs单独三维培养组.结论:HGFs和HUVECs共培养能够引导和促进HUVECs出芽及迁移.
Abstract
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.
关键词
3D生物打印/成血管/组织工程/人脐静脉内皮细胞/人牙龈成纤维细胞Key words
3D bioprinting/vascularization/tissue en-gineering/human umbilical vein endothelial cells/human gingival fibroblasts引用本文复制引用
基金项目
国家自然科学基金(82201071)
江苏省研究生科研与实践创新计划(KYCX23_2949)
出版年
2024
NETL
NSTL
万方数据