文献类型：期刊文献

英文题名：Fabrication of viable and functional pre-vascularized modular bone tissues by coculturing MSCs and HUVECs on microcarriers in spinner flasks

作者：Zhang, Songjie[1];Zhou, Min[1];Ye, Zhaoyang[1];Zhou, Yan[1];Tan, Wen-Song[1]

机构：[1]East China Univ Sci & Technol, Sch Bioengn, State Key Lab Bioreactor Engn, Shanghai, Peoples R China

年份：2017

卷号：12

期号：8

外文期刊名：BIOTECHNOLOGY JOURNAL

收录：;EI(收录号：20172703898595);WOS:【SCI-EXPANDED(收录号:WOS:000407059900011)】；

基金：This research was supported by the National Natural Science Foundation of China (Grant No. 31170951 and 81671841), the Natural Science Foundation of Shanghai (Grant No. 16ZR1408700) and Basic Research Key Program Project of Commission of Science and Technology of Shanghai (16JC1400203).

语种：英文

外文关键词：Bone tissue engineering; Endothelial cells; Mesenchymal stem cells; Microcarriers; Vascularization

摘要：Slow vascularization often impedes the viability and function of engineered bone replacements. Prevascularization is a promising way to solve this problem. In this study, a new process was developed by integrating microcarrier culture and coculture to fabricate pre-vascularized bone microtissues with mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs). Initially, coculture medium and cell ratio between MSCs and HUVECs were optimized in tissue culture plates concerning cell proliferation, osteogenesis and angiogenesis. Subsequently, cells were seeded onto CultiSpher S microcarriers in spinner flasks and subjected to a two-stage (proliferative-osteogenic) culture process for four weeks. Both cells proliferated and functioned well in chosen medium and a 1 : 1 ratio between MSCs and HUVECs was chosen for better angiogenesis. After four weeks of culture in spinner flasks, the microtissues were formed with high cellularity, evenly distributed cells and tube formation ability. While coculture with HUVECs exerted an inhibitory effect on osteogenic differentiation of MSCs, with downregulated alkaline phosphatase activity, mineralization and gene expression of COLI, RUNX2 and OCN, this could be attenuated by employing a delayed seeding strategy of HUVECs against MSCs during the microtissue fabrication process. Conclusion: Collectively, this work established an effective method to fabricate pre-vascularized bone microtissues, which would lay a solid foundation for subsequent development of vascularized tissue grafts for bone regeneration.