高表达Ang1基因的间充质干细胞株的建立及其肺内转化研究演示文稿.ppt

This is the Icon of the plasmid contained Ang1 pLV，pCMV-dR8.74 and pMD2G co-transfer MSCs in passage 1,passage 2, passage 3 and passage 4 light jugular [dZˉgjulE] 颈的,颈静脉的 高表达Ang1基因的间充质干细胞的建立及其肺内转化研究 徐金富 瞿介明 李惠萍等 同济大学附属上海市肺科医院 复旦大学附属华东医院 High promise for angiopoietin-1(Ang1) in lung injury intervention Ang1 decreased in LPS induced lung injury anti-inflammation: inhibits endothelial permeability, neutrophil adherence and IL-8 production Anti-apoptosis role to endothelial cells Thurston G, Rudge JS, Ioffe E, et al. Angiopoietin-1 protects the adult vasculature against plasma leakage. Nat Med. 2000, 6: 460-463.Daly C, Wong V, Burova E, et al. Angiopoietin-1 modulates endothelial cell function and gene expression via the transcription factor FKHR (FOXO1). Gene Dev. 2004, 18(9): 1060-1071.Pizurki L, Zhou Z, Glynos K, et al. Angiopoietin-1 inhibits endothelial permeability, neutrophil adherence and IL-8 production. Brit J Pharmacol. 2003, 139(2): 329-336. Mesenchymal stem cells (MSC) No ethical difficulties harvest MSC from the individual to be treated No rejection problems Self migrate to where the injury is located Rojas M, Xu J, Woods CR, et al. Bone marrow-derived mesenchymal stem cells in repair of the injured lung. Am J Respir Cell Mol Biol 2005; 33: 145–152Robert JD，Annemarie BM．mesenchymal stem cells：biology and potential clinical uses. Experimental Hematology, 2000, 28(8):87 Our hypothesis +Ang1 MSCs By Lentiviral vector MSCs transduced with Ang1 Mice LPS induced lung injury In vivo intervention Methods and results pCR2.1-Ang1 : Prof. Federico Bussolino in Institute for cancer research and treatment, Italy Sequencing results: 100% coincide with Ang1 cDNA sequence Electrophoresis after cutting by restriction enzyme pLV pCMV-dR8.74 pMD2G In vitro Lentivirus-mediated Ang1/eGFP transfer into MSCs Isolation, purification and expansion of MSCs Bone marrow was harvested from 6 weeks old C57BL/6 by flushing the femurs and tibias Sep