In vitro study of matrix metalloproteinase/tissue inhibitor of metalloproteinase production by mesenchymal stromal cells in response to inflammatory cytokines: the role of their migration in injured tissues.
OBJECTIVE
The transmigratory capacity of bone marrow (BM) mesenchymal stromal cells (MSC) through the endothelial cell barrier into various tissues and their differentiation potential makes them ideal candidates for cell therapy. Nevertheless, the mechanisms and agents promoting their migration are not fully understood. We evaluated the effects of several inflammatory cytokines on the migration of BM MSC and matrix metalloproteinase (MMP)/tissue inhibitor of metalloproteinase (TIMP) production.


METHODS
The migratory potential of BM MSC was evaluated using a Boyden chamber coated with Matrigel in the presence and absence of stromal cell-derived (SDF)-1alpha, platelet-derived growth factor (PDGF)bb, insulin-like growth factor (IGF)-I and interleukin (IL)-6. The ability of inflammatory cytokines to induce MSC migration was tested in presence of their respective Ab or blocking peptide. We used immunofluorescence to check the expression of cytokine receptors, and MMP/TIMP production was analyzed at the protein (human cytokine array, enzyme-linked immunosorbent assay (ELISA), gelatine zymography and Western blot) and mRNA quantitative real-time polymerase chain reaction (qRT-PCR) levels.


RESULTS
We have demonstrated that inflammatory cytokines promote the migratory capacity of BM MSC according to the expression of their respective receptors. Higher migration through Matrigel was observed in response to IL-6 and PDGFbb. qRT-PCR and cytokine array revealed that migration was the result of the variable level of MMP/TIMP in response to inflammatory stimuli.


CONCLUSIONS
Our observations suggest that chemokines and cytokines involved in the regulation of the immunity or inflammatory process promote the migration of MSC into BM or damaged tissues. One of the mechanisms used by MSC to promote their migration though the extracellular matrix is modulation of the production of MMP-1, MMP-2, MMP-13, TIMP-1 and TIMP-2.
Tondreau
Tatiana
T
Laboratory of Experimental Hematology, Institut Jules Bordet-ULB, Brussels, Belgium.
Meuleman
Nathalie
N
Stamatopoulos
Basile
B
De Bruyn
Cecile
C
Delforge
Alain
A
Dejeneffe
Marielle
M
Martiat
Philippe
P
Bron
Dominique
D
Lagneaux
Laurence
L
D001854:Bone Marrow Cells / Q000166:cytology; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D018925:Chemokines / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism; D016207:Cytokines / Q000494:pharmacology*; D004338:Drug Combinations; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D007249:Inflammation / Q000473:pathology; D018836:Inflammation Mediators / Q000494:pharmacology*; D007797:Laminin / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000096:biosynthesis* / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010641:Phenotype; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019707:Receptors, Chemokine / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D019714:Tissue Inhibitor of Metalloproteinases / Q000096:biosynthesis* / Q000235:genetics
D018925:Chemokines; D016207:Cytokines; D004338:Drug Combinations; D018836:Inflammation Mediators; D007797:Laminin; D011509:Proteoglycans; D012333:RNA, Messenger; D019707:Receptors, Chemokine; D019714:Tissue Inhibitor of Metalloproteinases; C059485:matrigel; D003094:Collagen; D020782:Matrix Metalloproteinases