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Original Article |
1 Joint Program in Transfusion Medicine, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts
2 Divisions of Allergy-Inflammation and Infectious Diseases at the Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts
* To whom correspondence should be addressed. E-mail: leslie.silberstein{at}childrens.harvard.edu.
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Abstract |
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Stromal cells isolated from bone marrow (BMSC) often referred to as mesenchymal stem cells, are currently under investigation for a variety of therapeutic applications. However, limited data are available regarding receptors that can influence their homing to and positioning within the bone marrow. In the present study we found that second passage BMSC express a unique set of chemokine receptors; three CC chemokine receptors (CCR1, CCR7 and CCR9) and three CXC chemokine receptors (CXCR4, CXCR5 and CXCR6). BMSC cultured in serum free medium secrete several chemokine ligands (CCL2, CCL4, CCL5, CCL20, CXCL12, CXCL8 and CX3CL1). The surface expressed chemokine receptors were functional by several criteria. Stimulation of BMSC with chemokine ligands triggers phosphorylation of the MAPK (e.g. ERK1 and ERK2) and focal adhesion kinase (FAK) signaling pathways. In addition, CXCL12 selectively activates STAT 5 whereas CCL5 activates STAT 1. In cell biologic assays, all of the chemokines tested stimulate chemotaxis of BMSCs and CXCL12 induces cytoskeleton F-actin polymerization. Studies of culture expanded BMSC, e.g. 12-16 passages, indicate loss of surface expression of all chemokine receptors and lack of chemotactic response to chemokines. The loss in chemokine receptor expression is accompanied by a decrease in expression of adhesion molecules (ICAM-1, ICAM-2, VCAM-1) and CD157 while expression of CD90 and CD105 is maintained. The change in BMSC phenotype is associated with slowing of cell growth and increased spontaneous apoptosis. These findings suggest that several chemokine axes may operate in bone marrow stromal cell biology and may be important parameters in the validation of cultured BMSC intended for cell therapy.
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