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Stem Cells International
Volume 2012 (2012), Article ID 920581, 10 pages
Research Article

Multipotent Mesenchymal Stromal Stem Cell Expansion by Plating Whole Bone Marrow at a Low Cellular Density: A More Advantageous Method for Clinical Use

1Stem Cell Transplantation and Cellular Therapy Unit, Pediatric Onco-Hematology Department, Regina Margherita Children's Hospital, 10126 Turin, Italy
2Department of Pediatrics, University of Turin, Piazza Polonia 94, 10126 Turin, Italy
3Molecular Biotechnology Center, University of Turin, Polonia 94, 10126 Turin, Italy

Received 13 May 2011; Accepted 1 August 2011

Academic Editor: Peter Schlenke

Copyright © 2012 Katia Mareschi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Mesenchymal stem cells (MSCs) are a promising source for cell therapy due to their pluripotency and immunomodulant proprieties. As the identification of “optimal” conditions is important to identify a standard procedure for clinical use. Percoll, Ficoll and whole bone marrow directly plated were tested from the same sample as separation methods. The cells were seeded at the following densities: 100 000, 10 000, 1000, 100, 10 cells/cm2. After reaching confluence, the cells were detached, pooled and re-plated at 1000, 500, 100, and 10 cells/cm2. Statistical analyses were performed. Cumulative Population Doublings (PD) did not show significant differences for the separation methods and seeding densities but only for the plating density. Some small quantity samples plated in T25 flasks at plating densities of 10 and 100 cells/cm2 did not produce any expansion. However, directly plated whole bone marrow resulted in a more advantageous method in terms of CFU-F number, cellular growth and minimal manipulation. No differences were observed in terms of gross morphology, differentiation potential or immunophenotype. These data suggest that plating whole bone marrow at a low cellular density may represent a good procedure for MSC expansion for clinical use.