title: A porous membrane-mediated isolation of mesenchymal stem cells from human embryonic stem cells.
abstract: Pluripotent human embryonic stem cells (hESCs) acquire mesenchymal characteristics during the epithelial-mesenchymal transition (EMT) process. Here, we report a simple and an efficient isolation method for mesenchymal stem cells (MSCs) from hESCs undergoing EMT using a commercialized porous membrane transwell culture insert. Suspension culture of hESC colonies results in the formation of embryoid bodies, which adhered on the upper compartment of 8 μm porous membrane in the presence of EMG2-MV media. The population migrating through the permeable membrane to the lower compartment not only exhibited EMT markers but also expressed high levels of a panel of typical MSC surface antigen markers, and demonstrated multipotent differentiation capability. In addition, they have a prolonged proliferation capacity without characteristics and chromosomal changes. Furthermore, the isolated MSCs significantly enhanced cardiac functions in a rat model of myocardial infarction (MI) as measured by the left ventricle wall thickness (MI control, 32.9%±3.2% vs. hESCs-MSCs, 38.7%±2.4%), scar length (MI control, 46.1%±2.5% vs. hESCs-MSCs, 41.8%±1.3%), fibrosis area (MI control, 34.3%±1.6% vs. hESCs-MSCs, 28.9%±3.5%), and capillary density. Our findings demonstrate an ease with which hESCs-MSCs can be effectively isolated using the porous membrane, which overcomes the lack of availability of MSCs for therapeutic applications in various diseased animal models.
authors.affiliation: 1 Department of Stem Cell Biology, School of Medicine, Konkuk University , Seoul, Republic of Korea.
authors.forename: Ki-Sung
authors.initials: KS
authors.lastname: Hong
authors.forename: Daekyeong
authors.initials: D
authors.lastname: Bae
authors.forename: Youngsok
authors.initials: Y
authors.lastname: Choi
authors.forename: Sun-Woong
authors.initials: SW
authors.lastname: Kang
authors.forename: Sung-Hwan
authors.initials: SH
authors.lastname: Moon
authors.forename: Hoon Taek
authors.initials: HT
authors.lastname: Lee
authors.forename: Hyung-Min
authors.initials: HM
authors.lastname: Chung
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008567:Membranes, Artificial; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008819:Mice, Nude; D009203:Myocardial Infarction / Q000628:therapy; D016062:Porosity; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D008567:Membranes, Artificial