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Embryonic Stem Cells |
1 Department of Surgery, National University of Singapore, Singapore, Singapore; Genome Institute of Singapore, Singapore, Singapore
2 Genome Institute of Singapore, Singapore, Singapore
3 Department of Pathology, National University of Singapore, Singapore, Singapore
4 Genome Institute of Singapore, Singapore, Singapore; Department of Orthopaedic Surgery, National University of Singapore, Singapore, Singapore
5 Department of Surgery, National University of Singapore, Singapore, Singapore
6 Department of Orthopaedic Surgery, National University of Singapore, Singapore, Singapore
7 Genome Institute of Singapore, Singapore, Singapore; Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts
8 Genome Institute of Singapore, Singapore, Singapore; Department of Biochemistry, National University of Singapore, Singapore, Singapore
* To whom correspondence should be addressed. E-mail: limsk{at}gis.a-star.edu.sg.
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Abstract |
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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 FACS 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 (r2>0.90). We have developed a protocol to reproducibly generate clinically compliant and identical hESC-MSC cultures.
Key Words. Human embryonic stem cells, mesenchymal stem cells, cell surface markers, adipogenesis, Chondrogenesis, Osteoprogenitor, Selectable marker, Somatic stem cells
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