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Characterization of bone marrow derived mesenchymal stem cells in suspension

Kentaro Akiyama12, Yong-Ouk You1, Takayoshi Yamaza13, Chider Chen1, Liang Tang4, Yan Jin4, Xiao-Dong Chen5, Stan Gronthos6 and Songtao Shi1*

Author Affiliations

1 Center for Craniofacial Molecular Biology, University of Southern California, 2250 Alcazar Street, CSA 103, Los Angeles, CA 90033, USA

2 Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Science, 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8525, Japan

3 Department of Molecular Cell Biology and Oral Anatomy, Kyushu University Graduate School of Dental Science, Fukuoka 812-8582, Japan

4 Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi'an, Shanxi, China

5 Division of Research, Department of Comprehensive Dentistry, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, Texas 78229-3900, USA

6 Mesenchymal Stem Cell Group, Department of Haematology, Institute of Medical and Veterinary Science/ Hanson Institute, Adelaide 5000, South Australia, Australia

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Stem Cell Research & Therapy 2012, 3:40  doi:10.1186/scrt131

Published: 19 October 2012



Bone marrow mesenchymal stem cells (BMMSCs) are a heterogeneous population of postnatal precursor cells with the capacity of adhering to culture dishes generating colony-forming unit-fibroblasts (CFU-F). Here we identify a new subset of BMMSCs that fail to adhere to plastic culture dishes and remain in culture suspension (S-BMMSCs).


To catch S-BMMSCs, we used BMMSCs-produced extracellular cell matrix (ECM)-coated dishes. Isolated S-BMMSCs were analyzed by in vitro stem cell analysis approaches, including flow cytometry, inductive multiple differentiation, western blot and in vivo implantation to assess the bone regeneration ability of S-BMMSCs. Furthermore, we performed systemic S-BMMSCs transplantation to treat systemic lupus erythematosus (SLE)-like MRL/lpr mice.


S-BMMSCs are capable of adhering to ECM-coated dishes and showing mesenchymal stem cell characteristics with distinction from hematopoietic cells as evidenced by co-expression of CD73 or Oct-4 with CD34, forming a single colony cluster on ECM, and failure to differentiate into hematopoietic cell lineage. Moreover, we found that culture-expanded S-BMMSCs exhibited significantly increased immunomodulatory capacities in vitro and an efficacious treatment for SLE-like MRL/lpr mice by rebalancing regulatory T cells (Tregs) and T helper 17 cells (Th17) through high NO production.


These data suggest that it is feasible to improve immunotherapy by identifying a new subset BMMSCs.