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TISSUE-SPECIFIC STEM CELLS

Human Multipotent Stromal Cells Undergo Sharp Transition from Division to Development in Culture

Center for Gene Therapy, Tulane University Health Sciences Center, New Orleans, Louisiana, USA

Key Words. Multipotent stromal cells • Microarray • Mesenchymal stem cells • Transcriptome • Transplantation

Correspondence: Correspondence: Darwin J. Prockop, M.D., Ph.D., Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, Louisiana 70112, USA. Telephone: 504-988-7711; Fax: 504-988-7710; e-mail: dprocko{at}tulane.edu

Received on July 3, 2007; accepted for publication on September 21, 2007.

First published online in STEM CELLS EXPRESS  October 4, 2007.


Human mesenchymal stem cells, or multipotent stromal cells (MSCs), are of interest for clinical therapy, in part because of their capacity for proliferation and differentiation. However, results from clinical trials and in vitro models have been variable, possibly because of MSC heterogeneity and a lack of standardization between MSC in vitro expansion protocols. Here we defined changes in MSCs during expansion in vitro. In low-density cultures, MSCs expand through distinct lag, exponential growth, and stationary phases. We assayed cultures of passage 2 human MSCs from three donors at low density (50 cells per cm²) at approximately 5% confluence on day 2 and after the cultures had expanded to approximately 70% confluence on day 7. On day 2, genes involved in cell division were upregulated. On day 7, genes for cell development were upregulated. The variations among three donors were less than the variation within the expansion of MSCs from a single donor. The microarray data for selected genes were confirmed by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and FACScan analysis. Approximately 50% of cells at day 2 were in S-phase compared with 10% at day 7. The results demonstrated major differences in early and late stage cultures of MSCs that should be considered in using the cells in experiments and clinical applications.

Disclosure of potential conflicts of interest is found at the end of this article.

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