Ling Zhou1, Xuping Niu1, Jiannan Liang1, Junqin Li1, Jiao Li1, Yueai Cheng1, Yanfeng Meng1, Qiang Wang1, Xiaoli Yang1, Gang Wang2, Yu Shi3, Erle Dang2, Kaiming Zhang4. 1. Shanxi Key Laboratory of Stem Cell for Immunological Dermatosis, Institute of Dermatology, Taiyuan City Centre Hospital, No. 1 Dong San Dao Xiang, Jiefang Road, Taiyuan 030009, Shanxi Province, China(1). 2. Department of Dermatology, Xijing Hospital, Fourth Military Medical University, No. 15 Changle Road West, Xi'an, 710032, Shanxi Province, China. 3. Department of Haematology, Oncology and Tumor Immunology, Charité University Medicine Berlin, Campus Virchow Hospital, Berlin, Germany. 4. Shanxi Key Laboratory of Stem Cell for Immunological Dermatosis, Institute of Dermatology, Taiyuan City Centre Hospital, No. 1 Dong San Dao Xiang, Jiefang Road, Taiyuan 030009, Shanxi Province, China(1). Electronic address: zhangkaiming@sina.com.
Abstract
OBJECTIVE: To directionally-differentiate dermis-derived mesenchymal stem cells (DMSCs) into vascular endothelial cells (VECs) in vitro, providing an experimental basis for studies on the pathogenesis and treatment of vascular diseases. METHODS: After separation by adherent culture, VEC line supernatant, vascular endothelial growth factor (VEGF), bone morphogenetic protein-4 and hypoxia were used for the differentiation of VECs from DMSCs. The cell type was authenticated by flow cytometry, matrigel angiogenesis assay in vitro, and immunofluorescent staining during differentiation. The VEGF concentration was investigated by enzyme-linked immunosorbent assay. RESULTS: After 28 days of differentiation, the cell surface marker CD31 was significantly positive (80%-90%) by flow cytometry in the VEC line-conditioned culture, which was significantly higher than in the other groups. Differentiated DMSCs had the ability to ingest Dil-ac-LDL and vascularize in the conditioned culture, but not in the other groups. In the VEC line supernatant, the concentration of VEGF was very low. The VEGF concentration changed along with the differentiation into VECs in the medium of the conditioned culture group. CONCLUSION: VEC line supernatant can induce the differentiation of DMSCs into VECs, possibly through the pathway except VEGF.
OBJECTIVE: To directionally-differentiate dermis-derived mesenchymal stem cells (DMSCs) into vascular endothelial cells (VECs) in vitro, providing an experimental basis for studies on the pathogenesis and treatment of vascular diseases. METHODS: After separation by adherent culture, VEC line supernatant, vascular endothelial growth factor (VEGF), bone morphogenetic protein-4 and hypoxia were used for the differentiation of VECs from DMSCs. The cell type was authenticated by flow cytometry, matrigel angiogenesis assay in vitro, and immunofluorescent staining during differentiation. The VEGF concentration was investigated by enzyme-linked immunosorbent assay. RESULTS: After 28 days of differentiation, the cell surface marker CD31 was significantly positive (80%-90%) by flow cytometry in the VEC line-conditioned culture, which was significantly higher than in the other groups. Differentiated DMSCs had the ability to ingest Dil-ac-LDL and vascularize in the conditioned culture, but not in the other groups. In the VEC line supernatant, the concentration of VEGF was very low. The VEGF concentration changed along with the differentiation into VECs in the medium of the conditioned culture group. CONCLUSION: VEC line supernatant can induce the differentiation of DMSCs into VECs, possibly through the pathway except VEGF.