배지 성분에 따른 인간 지방조직기원 CD146 양성 혈관내피세포의 증식 및 기능의 평가

Abstract

Active blood supply is an essential pre-requisite for tissue to survive and integrate with existing host tissues. Without proper blood supply, seeded osteoprogenitor cells in the scaffold would be subject to necrosis due to inappropriate metabolites. Ineffective vascularization often restrains new bone formation. We cultured adipose tissue-derived CD146 positive endothelial cells using adipose tissue harvested form buccal fat pad in the previous study. Using polydioxanone/pluronic F127 scaffold, we would like to improve the osteogenesis of engineered graft by pre-seeding adipose tissue-derived endothelial cells which precipitated the vascularization. Prior to the formation of vascularized polydioxanone/pluronic F127 scaffold, this study focused on proliferation and function of the adipose tissue-derived endothelial cells according to culture mediums. Adipose tissue-derived CD146 positive endothelial cells were cultured in previous technique. These cells were further cultured according to different culture mediums (DMEM culture medium with or without osteogenic inductive agents and EGM-2 culture medium with or without osteogenic inductive agents). After 24 hours of culture, among the culture medium using DMEM, the proliferation of adipose tissue-derived endothelial cells were promoted in DMEM medium without osteogenic inductive agents than in DMEM with osteogenic inductive agents. However, 72 hours of culture, the proliferation of the cells were promoted in DMEM medium with osteogenic inductive agents than in DMEM without osteogenic inductive agents. Among the culture medium using EGM-2, the proliferation of the cells were promoted in EGM-2 medium without osteogenic inductive agents than in EGM-2 medium with osteogenic inductive agents. However, 72 hours of culture, the proliferation of the cells were promoted in EGM-2 medium with osteogenic inductive agents than in EGM-2 medium without osteogenic inductive agents. And then tube formation was examined about the function of adipose tissue-derived endothelial cells. These adipose tissue-derived cells formed tube-like structures on Matrigel in EGM-2 culture medium with or without osteogenic inductive agents. However, the cells did not form tube-like structures on Matrigel in DMEM medium with or without osteogenic inductive agents. These results suggest that the proliferation and function of the adipose tissue-derived CD146 positive endothelial cells could be maintained in EGM-2 with osteogenic inductive agents. However, the function of the cells could not be maintained in DMEM with or without osteogenic inductive agents.