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

Harkness, Linda, Mahmood, Amer, Ditzel, Nicholas, Abdallah, Basem M., Nygaard, Jens V. and Kassem, Moustapha (2011) Selective isolation and differentiation of a stromal population of human embryonic stem cells with osteogenic potential. Bone, 48 2: 231-241. doi:10.1016/j.bone.2010.09.023


Author Harkness, Linda
Mahmood, Amer
Ditzel, Nicholas
Abdallah, Basem M.
Nygaard, Jens V.
Kassem, Moustapha
Title Selective isolation and differentiation of a stromal population of human embryonic stem cells with osteogenic potential
Journal name Bone   Check publisher's open access policy
ISSN 8756-3282
1873-2763
Publication date 2011-02-01
Sub-type Article (original research)
DOI 10.1016/j.bone.2010.09.023
Open Access Status Not Open Access
Volume 48
Issue 2
Start page 231
End page 241
Total pages 11
Place of publication Philadelphia, PA, United States
Publisher Elsevier
Language eng
Abstract 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 (100μg/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 10weeks, 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.
Keyword Human embryonic stem cells
Hyaluronic acid
In vivo bone formation
Osteogenic differentiation
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Wed, 23 Sep 2015, 15:33:39 EST by Linda Harkness on behalf of Scholarly Communication and Digitisation Service