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First published online May 25, 2006
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2005-0591v1
24/9/2052    most recent
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Submitted on November 25, 2005
Accepted on May 16, 2006

Technology Development

Bioreactor Expansion of Human Adult Bone Marrow-derived Mesenchymal Stem Cells (MSCs)

Xi Chen 1, Hai-bo Xu 2, Chao Wan 1, Mervyn McCaigue 1, Gang Li 1*

1 Department of Orthopaedic Surgery, Center for Cancer Research and Cell Biology, Queen's University Belfast, Belfast, United Kingdom
2 Department of Surgery, Center for Cancer Research and Cell Biology, Queen's University Belfast, Belfast, United Kingdom

* To whom correspondence should be addressed. E-mail: g.li{at}qub.ac.uk.


  Abstract

Supplementation of mesenchymal stem cells (MSCs) during hematopoietic stem cell transplantation (HSCT) alleviates complications such as graft-versus-host disease, leading to a speedy recovery of hematopoiesis. To meet such clinical demand, a fast MSCs expansion method is required. In the present study, we examined the feasibility of expanding MSCs from the isolated bone marrow mononuclear cells using a rotary bioreactor system. The cells were cultured in a rotary bioreactor with MyelocultTM medium containing a combination of supplementary factors, including stem cell factor (SCF), interleukin 3 and 6 (IL-3, IL-6). After 8 days of culture, total cell numbers, Stro-1+CD44+CD34- MSCs and CD34+CD44+Stro-1- HSCs were increased 9, 29, and 8 folds respectively. Colony forming efficiency-fibroblast per day (CFE-F/day) of the bioreactor-treated cells was 1.44-fold higher than that of the cells without bioreactor treatment. The bioreactor-expanded MSCs showed expression of primitive MSCs markers endoglin (SH2) and vimentin, whereas markers associated with lineage differentiation including osteocalcin (osteogenesis), Type II collagen (chondrogenesis) and C/EBP{alpha} (adipogenesis) were not detected. Upon induction, the bioreactor-expanded MSCs were able to differentiate into osteoblasts, chondrocytes and adipocytes. Taken together, we conclude that the rotary bioreactor with the modified MyelocultTM medium reported in this study may be used to rapidly expand MSCs.

Key Words. mesenchymal stem cells, hematopoietic stem cells, bioreactor, differentiation




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