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First published online October 19, 2006
Stem Cells Vol. 25 No. 2 February 2007, pp. 425 -436
doi:10.1634/stemcells.2006-0420; www.StemCells.com
© 2007 AlphaMed Press

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EMBRYONIC STEM CELLS

Derivation of Clinically Compliant MSCs from CD105+, CD24– Differentiated Human ESCs

Qizhou Liana,b, Elias Lyea,b, Keng Suan Yeob, Eileen Khia Way Tanb, Manuel Salto-Tellezc, Tong Ming Liub,d, Nallasivam Palanisamyb, Reida Menshawe El Oakleya, Eng Hin Leed, Bing Limb,e, Sai-Kiang Limb,f

aDepartment of Surgery, National University of Singapore, Singapore;
bGenome Institute of Singapore, Singapore;
cDepartment of Pathology, National University of Singapore, Singapore;
dDepartment of Orthopaedic Surgery, National University of Singapore, Singapore;
eBeth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA;
fDepartment of Biochemistry, National University of Singapore, Singapore

Key Words. Human embryonic stem cells • Mesenchymal stem cells • Cell surface markers • Adipogenesis • Chondrogenesis Osteoprogenitor • Selectable marker • Somatic stem cells

Correspondence: Sai Kiang Lim, Ph.D., Stem Cell and Developmental Biology Group I, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672. Telephone: 6478-8146; Fax: 6478-9005; e-mail: limsk{at}gis.a-star.edu.sg

Received on July 12, 2006; accepted for publication on October 11, 2006.

First published online in STEM CELLS EXPRESS  October 19, 2006.


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 (r2 > .90). We have developed a protocol to reproducibly generate clinically compliant and identical hESC-MSC cultures.




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