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Migration of culture-expanded human mesenchymal stem cells through bone marrow endothelium is regulated by matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3

From the Stem Cell Laboratory (ADB, IVB, JDW, IVR) and Laboratory of Hematology-Immunology, Academic Hospital Vrije Universiteit Brussel (VUB), Brussels, Belgium (MB, MDW); MicroArray Facility, Flanders Interuniversitary Institute for Biotechnology (VIB), Leuven, Belgium (PVH).

Correspondence: Ivan Van Riet, Stem Cell Laboratory, Academic Hospital Vrije Universiteit Brussel (VUB) Research group, Hematology-Immunology VUB, Laarbeeklaan 105, B-1090 Brussels, Belgium. E-mail: ivan.vanriet{at}az.vub.ac.be

Background and Objectives: Mesenchymal stem cells (MSC) are adult stem cells that can be expanded many fold in vitro and have the therapeutic potential to restore the bone marrow microenvironment and support hematopoietic recovery after myeloablative conditioning for hematopoietic stem cell transplantation. Successful homing to the target tissue, such as bone marrow, implies that MSC are able to extravasate after systemic administration. However, the extravasation capacity of MSC and the underlying mechanisms are poorly understood to date. We studied in vitro the capacity of MSC to migrate through bone marrow endothelium.

Design and Methods: In vitro invasion and transendothelial migration assays were performed. The expression of matrix metalloproteinase (MMP) was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Migration of cells cultured at high or low confluence was compared and differential gene expression in these conditions was analyzed with microarray and real-time RT-PCR. The functional involvement in MSC migration was assessed using neutralizing anti-MMP-2 antibody, MMP-2 short interfering RNA or recombinant tissue inhibitor of metalloproteinase (TIMP-3).

Results: We demonstrated that MSC can invade reconstituted basement membrane and that bone marrow endothelial cells stimulate this process. We also showed that the transendothelial migration of MSC is at least partially regulated by MMP-2. High culture confluence was found to increase production of the natural MMP-inhibitor TIMP-3 and decrease transendothelial migration of MSC.

Interpretation and Conclusions: We show that MSC have the potential to migrate through bone marrow endothelium and that this process involves MMP-2. Moreover, the migration of MSC is significantly influenced by the level of culture confluence. Increased culture confluence impairs migration and is related to an upregulation of TIMP-3. The therapeutic use of MSC would benefit from a selection of culture conditions that allow optimal extravasation of these cells.

Key words: mesenchymal stem cell, migration, culture confluence, MMP-2, TIMP-3.