Mesenchymal stem cells, used as bait, disclose tissue binding sites: a tool in the search for the niche?
We developed an ex vivo approach characterizing renal mesenchymal stem cell (MSC) adhesion to kidney sections. Specificity of MSC adhesion was confirmed by demonstrating a) 3T3 cells displayed 10-fold lower adhesion, and b) MSC adhesion was CXCR4/stromal-derived factor-1 (SDF-1)-dependent. MSC adhesion was asymmetrical, with postischemic sections exhibiting more than twofold higher adhesion than controls, and showed preference to perivascular areas. Pretreating kidney sections with cyclic arginine-glycine-aspartic acid peptide resulted in increased MSC adhesion (by displacing resident cells), whereas blockade of CXCR4 with AMD3100 and inhibition of alpha4beta1(VLA4) integrin or vascular cellular adhesion molecule-1, reduced adhesion. The difference between adhered cells under cyclic arginine-glycine-aspartic acid peptide-treated and control conditions reflected prior occupancy of binding sites with endogenous cells. The AMD3100-inhibitable fraction of adhesion reflected CXCR4-dependent adhesion, whereas maximal adhesion was interpreted as kidney MSC-lodging capacity. MSC obtained from mice overexpressing caveolin-1 exhibited more robust adhesion than those obtained from knockout animals, consistent with CXCR4 dimerization in caveolae. These data demonstrate a) CXCR4/SDF-1-dependent adhesion increases in ischemia; b) CXCR4/SDF-1 activation is dependent on MSC surface caveolin-1; and c) occupancy of MSC binding sites is decreased, while d) capacity of MSC binding sites is expanded in postischemic kidneys. In conclusion, we developed a cell-bait strategy to unmask renal stem cell binding sites, which may potentially shed light on the MSC niche(s) and its characteristics.
Ratliff
Brian B
BB
Department of Medicine, New York Medical College, Renal Research Institute, Valhalla, NY 10595, USA.
Singh
Nandita
N
Yasuda
Kaoru
K
Park
Hyeong-Cheon
HC
Addabbo
Francesco
F
Ghaly
Tammer
T
Rajdev
Maharshi
M
Jasmin
Jean-Francois
JF
Plotkin
Matthew
M
Lisanti
Michael P
MP
Goligorsky
Michael S
MS
D016475:3T3 Cells; D000818:Animals; D001665:Binding Sites; D051242:Caveolin 1 / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D016023:Integrins / Q000378:metabolism; D007668:Kidney / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D019718:Receptors, CXCR4 / Q000378:metabolism; D055153:Stem Cell Niche
D051242:Caveolin 1; D054377:Chemokine CXCL12; C515256:Cxcl12 protein, mouse; D016023:Integrins; D019718:Receptors, CXCR4