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The derivation of osteogenic cells from human embryonic stem cells (hESC) has been hampered by the absence of easy and reproducible protocols. hESC grown in feeder-free conditions, often show a sub population of fibroblast-like, stromal cells growing between the colonies. Thus, we examined the possibility that these cells represent a population of stromal (mesenchymal) stem cells (hESC-stromal). Two in house derived hES cell lines (Odense3 and KMEB3) as well as an externally derived cell line (Hues8) were transitioned to feeder-free conditions. A sub population of fibroblast-like cells established between the hESC colonies were isolated by selective adherence to hyaluronic acid-coated plates (100 μg/ml) and were characterized using a combination of FACS analysis and staining. The cells were CD44 + , CD29 + , CD73 + , CD166 + , CD146 + , and CD105 + ; and, Oct4 − , CD34 − , CD45 − and CXCR4 − . When cultured in osteogenic differentiation media, up regulation of osteoblastic lineage markers ( DLX5, MSX2, RUNX2, SPARC, ALP, COL1a1, BGLAP, IBSP, DCN, LOX-L4 ) and production of in vitro mineralized matrix was detected. hESC-stromal cells loaded on a carrier and implanted either subcutaneously or in a critical size calvarial defect in immune deficient mice for 10 weeks, resulted in new bone formation and partial repair of the calvarial defect. In conclusion, hESC-stromal can be isolated from hESC cultures and represent a good source for obtaining cells with osteogenic differentiation potential suitable for regenerative medicine protocols.

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Selective isolation and differentiation of a stromal population of human embryonic stem cells with osteogenic potential

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  • Publisher Elsevier
  • Copyright Copyright © 2010 Elsevier Inc.
  • ISSN 8756-3282
  • D.O.I. 10.1016/j.bone.2010.09.023
  • Publisher site Get PDF  

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