title: Umbilical cord-derived mesenchymal stem cells isolated by a novel explantation technique can differentiate into functional endothelial cells and promote revascularization.
abstract: Stem cells transplantation holds great promise for the treatment of ischemic diseases through functional revascularization. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are also an ideal candidate for cell-based bioengineering. Herein, we report on the development of a simple and effective protocol to isolate UC-MSCs, and confirm their endothelial potential both in vitro and in vivo. UC-MSCs were isolated by a novel explantation technique and induced to differentiate into endothelial-like cells. Then UC-MSCs were transplanted into ischemic mouse model and cultured on 3D gel/MMT-CS composite scaffolds. Morphological and proliferation assessments show that sufficient UC-MSCs can be generated during a relatively short culture period with explantation technique. Increased expression of endothelial-specific markers (KDR and vWF), and functional markers (ac-LDL uptake and UEA-1 binding), indicate that functional endothelial progenitor cells are induced after 9 days of in vitro culture. In an ischemic hindlimb mouse model, the ratio of ischemic/nonischemic limb perfusion 4 weeks after MSCs transplantation reached 0.84 +/- 0.09. The capillary density of this group was 2.57-fold greater than that of sham-injected mice (P < 0.05). Immunofluorescence and immunohistological analyses indicate that MSCs may act to salvage the ischemic tissue by incorporating into the local vasculature. In vitro, UC-MSCs were observed to incorporate into 3D gel/MMT-CS composite scaffolds, to secrete extracellular matrix, to remain viable, and to retain their proliferation capacity. In conclusion, UC-MSCs isolated by novel yet simple explantation technique are well suited for application in the development of novel stem cell-based revascularization therapies.
authors.affiliation: Institute of Hematology & Hospital of Blood Diseases, State Key Laboratory of Experimental Hematology, Chinese Academy of Medical Sciences & Peking Union of Medical College, Tianjin, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.forename: Hengxing
authors.initials: H
authors.lastname: Meng
authors.forename: Changhong
authors.initials: C
authors.lastname: Li
authors.forename: Mu
authors.initials: M
authors.lastname: Hao
authors.forename: Yafei
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yu
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.forename: Junling
authors.initials: J
authors.lastname: Han
authors.forename: Qiongli
authors.initials: Q
authors.lastname: Zhai
authors.forename: Lugui
authors.initials: L
authors.lastname: Qiu
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000502:physiology; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D006614:Hindlimb / Q000098:blood supply / Q000473:pathology; D006801:Humans; D007511:Ischemia / Q000473:pathology / Q000628:therapy; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010011:Osteocytes / Q000166:cytology / Q000502:physiology; D014470:Umbilical Cord / Q000166:cytology*