title: Derivation of clinically compliant MSCs from CD105+, CD24- differentiated human ESCs.
abstract: Adult tissue-derived mesenchymal stem cells (MSCs) have demonstrated therapeutic efficacy in treating diseases or repairing damaged tissues through mechanisms thought to be mediated by either cell replacement or secretion of paracrine factors. Characterized, self-renewing human ESCs could potentially be an invariable source of consistently uniform MSCs for therapeutic applications. Here we describe a clinically relevant and reproducible manner of generating identical batches of hESC-derived MSC (hESC-MSC) cultures that circumvents exposure to virus, mouse cells, or serum. Trypsinization and propagation of HuES9 or H1 hESCs in feeder- and serum-free selection media generated three polyclonal, karyotypically stable, and phenotypically MSC-like cultures that do not express pluripotency-associated markers but displayed MSC-like surface antigens and gene expression profile. They differentiate into adipocytes, osteocytes, and chondrocytes in vitro. Gene expression and fluorescence-activated cell sorter analysis identified CD105 and CD24 as highly expressed antigens on hESC-MSCs and hESCs, respectively. CD105+, CD24- monoclonal isolates have a typical MSC gene expression profiles and were identical to each other with a highly correlated gene expression profile (r(2) > .90). We have developed a protocol to reproducibly generate clinically compliant and identical hESC-MSC cultures.
authors.affiliation: Department of Surgery, National University of Singapore, Singapore.
authors.forename: Qizhou
authors.initials: Q
authors.lastname: Lian
authors.forename: Elias
authors.initials: E
authors.lastname: Lye
authors.forename: Keng
authors.initials: K
authors.lastname: Suan Yeo
authors.forename: Eileen
authors.initials: E
authors.lastname: Khia Way Tan
authors.forename: Manuel
authors.initials: M
authors.lastname: Salto-Tellez
authors.forename: Tong Ming
authors.initials: TM
authors.lastname: Liu
authors.forename: Nallasivam
authors.initials: N
authors.lastname: Palanisamy
authors.forename: Reida Menshawe
authors.initials: RM
authors.lastname: El Oakley
authors.forename: Eng Hin
authors.initials: EH
authors.lastname: Lee
authors.forename: Bing
authors.initials: B
authors.lastname: Lim
authors.forename: Sai-Kiang
authors.initials: SK
authors.lastname: Lim
mesh_terms: D050156:Adipogenesis; D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051927:CD24 Antigen / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D020219:Chondrogenesis; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D000071063:Endoglin; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D010012:Osteogenesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D033581:Stem Cell Transplantation
chemical_list: D015703:Antigens, CD; D051927:CD24 Antigen; C064206:ENG protein, human; D000071063:Endoglin; D012333:RNA, Messenger; D011956:Receptors, Cell Surface