内皮祖细胞的研究进展2008.pptVIP

Bone marrow Histopaque 1083 Methods Mononuclear cells (MNCs) were isolated from mouse bone marrow with Histopaque 1083 by density gradient centrifugation. Bone marrow Histopaque 1083 Resuspend in EGM2 Plate 5X106 cells/well on fibronectin-coated 6-well dish Methods MNCs were cultured in EGM2 with the supplement of VEGF, b-FGF, IGF-1 on fibronectin-coated 6-well plate for 2 days to remove mature endothelial cells and monocytes. Bone marrow Histopaque 1083 Resuspend in EGM2 Plate 5X106 cells/well on fibronectin-coated 6-well dish 2 days Collect no-adherent cells Plate 1X106 cells/well on fibronectin-coated 24-well dish Methods The non-adherent cells (which contain EPCs) are then harvested and plated on fibronectin-coated 24-well plates for 3 days. Bone marrow Histopaque 1083 Resuspend in EGM2 Plate 5X106 cells/well on fibronectin-coated 6-well dish 2 days Collect no-adherent cells Plate 1X106 cells/well on fibronectin-coated 24-well dish CD31, vWF and eNOS Methods 2/3 volume culture media was changed every 2 days. Immunofluorescence staining for CD31, vWF and eNOS were performed and were observed under fluromicroscope. Peripheral Blood Mononuclear Cells at Day 1 Peripheral Blood Mononuclear Cells at Day 2 * Department of Pathophysiology Hongmei TAN Nov, 2008 Definition They are circulating, bone marrow-derived cells that are functionally and phenotypically distinct from mature endothelial cells They can differentiate into endothelial cells in vitro, as assessed by expression profiles and functional characteristics They can contribute to in vivo vasculogenesis and / or vascular homeostasis 内皮祖细胞是能直接分化为血管内皮细胞的前体细胞 Embryonic EPC 胚外中胚层卵黄囊血岛 与造血干细胞存在共同的前体 Adult EPC Bone Marrow (3% BM-MNC) Peripheral Blood (0.2% MNC) Origin and differentiation of endothelial progenitor cells Hematopoietic stem cells CD133++ CD34+ CD45 CD14+ CD45+ CD45+ CD14++ CD45+ CD14+++ CD133+ KDR+ CD34+ CD14+ CD133+ Endothelial markers+ Endothelial markers+ EPCs E