G-CSF-mobilized peripheral blood mononuclear cells from diabetic patients augment neovascularization in ischemic limbs but with impaired capability.
BACKGROUND
Autologous transplantation of mobilized peripheral blood mononuclear cells (M-PBMNCs) is a novel approach to improve critical limb ischemia (CLI) in diabetes. However, endothelial progenitor cells (EPCs) from diabetes are dysfunctional and impaired in ischemia-induced neovascularization.


OBJECTIVE
This study aimed to confirm the compromised efficiency of diabetic M-PBMNCs in therapeutic neovascularization, and to determine the underlying mechanisms of this impairment.


METHODS
Diabetic M-PBMNCs from 17 diabetic patients or healthy controls, or phosphate-buffered saline (PBS) were injected into the ischemic limbs of streptozotocin-induced diabetic nude mice. The limb blood perfusion, ambulatory score, ischemia damage, capillary/fiber ratio, arteriole density, collateral vessel formation, and pericytes recruitment were evaluated between these three groups. Non-invasive real time image and histopathology were used to detect the in vivo role of transplanted M-PBMNCs. Proliferation and adhesion of EPCs were assayed. In vitro vascular network incorporation and matrigel plug assay were used to test the pro-neovascularization role of M-PBMNCs.


RESULTS
Transplantation of diabetic M-PBMNCs also improved neovascularization, but to a lesser extent from that observed with non-diabetic ones. This was associated with the impairment of diabetic M-PBMNCs capacity to differentiate into EPCs, to incorporate into vessel-like tubules in vitro, to participate in vascular-like structure formation in a subcutaneous matrigel plug, and to stimulate the recruitment of pericytes/smooth muscle cells. In addition, there was impairment in vasculogenesis, which was related to the reduced adhesion ability of EPCs from diabetic M-PBMNCs.


CONCLUSIONS
Diabetes reduced the capacity of M-PBMNCs to augment neovascularization in ischemia.
Zhou
B
B
State Key Laboratory of Experimental Hematology, National Research Center for Stem Cell Engineering and Technology, Institute of Hematology, Chinese Academy of Medical Sciences & Peking Union Medical College, 288 Nanjing Road, Tianjin 300020, China.
Bi
Y Y
YY
Han
Z B
ZB
Ren
H
H
Fang
Z H
ZH
Yu
X F
XF
Poon
M-C
MC
Han
Z C
ZC
D000368:Aged; D000369:Aged, 80 and over; D016022:Case-Control Studies; D017690:Cell Transplantation; D003920:Diabetes Mellitus / Q000097:blood / Q000503:physiopathology*; D005121:Extremities / Q000098:blood supply*; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage* / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007511:Ischemia / Q000628:therapy*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000187:drug effects*; D018919:Neovascularization, Physiologic
D016179:Granulocyte Colony-Stimulating Factor