Endothelial progenitor cells from human fetal aorta cure diabetic foot in a rat model.
OBJECTIVE
Recent evidence has suggested that circulating endothelial progenitor cells (EPCs) can repair the arterial endothelium during vascular injury. However, a reliable source of human EPCs is needed for therapeutic applications. In this study, we isolated human fetal aorta (HFA)-derived EPCs and analyzed the capacity of EPCs to differentiate into endothelial cells. In addition, because microvascular dysfunction is considered to be the major cause of diabetic foot (DF), we investigated whether transplantation of HFA-derived EPCs could treat DF in a rat model.


METHODS
EPCs were isolated from clinically aborted fetal aorta. RT-PCR, fluorescence-activated cell sorting, immunofluorescence, and an enzyme-linked immunosorbent assay were used to examine the expressions of CD133, CD34, CD31, Vascular Endothelial Growth Factor Receptor 2 (VEGFR2), von Willebrand Factor (vWF), and Endothelial Leukocyte Adhesion Molecule-1 (ELAM-1). Morphology and Dil-uptake were used to assess function of the EPCs. We then established a DF model by injecting microcarriers into the hind-limb arteries of Goto-Kakizaki rats and then transplanting the cultured EPCs into the ischemic hind limbs. Thermal infrared imaging, oxygen saturation apparatus, and laser Doppler perfusion imaging were used to monitor the progression of the disease. Immunohistochemistry was performed to examine the microvascular tissue formed by HFA-derived EPCs.


RESULTS
We found that CD133, CD34, and VEGFR2 were expressed by HFA-derived EPCs. After VEGF induction, CD133 expression was significantly decreased, but expression levels of vWF and ELAM-1 were markedly increased. Furthermore, tube formation and Dil-uptake were improved after VEGF induction. These observations suggest that EPCs could differentiate into endothelial cells. In the DF model, temperature, blood flow, and oxygen saturation were reduced but recovered to a nearly normal level following injection of the EPCs in the hind limb. Ischemic symptoms also improved. Injected EPCs were preferentially and durably engrafted into the blood vessels. In addition, anti-human CD31+-AMA+-vWF+ microvasculars were detected after transplantation of EPCs.


CONCLUSIONS
Early fetal aorta-derived EPCs possess strong self-renewal ability and can differentiate into endothelial cells. We demonstrated for the first time that transplanting HFA-derived EPCs could ameliorate DF prognosis in a rat model. These findings suggest that the transplantation of HFA-derived EPCs could serve as an innovative therapeutic strategy for managing DF.
Peking University China-Japan Friendship School of Clinical Medicine, Beijing, China.
Wan-Ni
WN
Zhao
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China.
Shi-Qing
SQ
Xu
Department of Neurosurgery, Peking University International Hospital, Beijing, China.
Jian-Feng
JF
Liang
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China.
Liang
L
Peng
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China.
Hong-Lin
HL
Liu
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China.
Zai
Z
Wang
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China.
Qing
Q
Fang
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China; Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Meng
M
Wang
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China; Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Wei-Qin
WQ
Yin
Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China. Electronic address: zwj-72@163.com.
Wen-Jian
WJ
Zhang
Peking University China-Japan Friendship School of Clinical Medicine, Beijing, China; Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing, China. Electronic address: lou.j@mail.com.
Jin-Ning
JN
Lou
D037881:Aborted Fetus / Q000166:cytology; D000818:Animals; D001011:Aorta / Q000166:cytology*; D002454:Cell Differentiation; D000066673:Cell Self Renewal; D017690:Cell Transplantation / Q000379:methods*; D017719:Diabetic Foot / Q000628:therapy*; D042783:Endothelial Cells; D066026:Endothelial Progenitor Cells / Q000166:cytology* / Q000502:physiology / Q000637:transplantation; D006801:Humans; D008297:Male; D018919:Neovascularization, Physiologic; D051381:Rats; D011919:Rats, Inbred Strains
Angiogenesis; Diabetic foot; Embryonic stem cells; Endothelial progenitor cells; Humans