Novel Cell-Free Strategy for Therapeutic Angiogenesis: In Vitro Generated Conditioned Medium Can Replace Progenitor Cell Transplantation

被引:183
作者
Di Santo, Stefano
Yang, Zijiang
von Ballmoos, Moritz Wyler
Voelzmann, Jan
Diehm, Nicolas
Baumgartner, Iris
Kalka, Christoph
机构
[1] Department of Vascular Medicine, Swiss Cardiovascular Center, University of Bern, Bern
关键词
D O I
10.1371/journal.pone.0005643
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Current evidence suggests that endothelial progenitor cells (EPC) contribute to ischemic tissue repair by both secretion of paracrine factors and incorporation into developing vessels. We tested the hypothesis that cell-free administration of paracrine factors secreted by cultured EPC may achieve an angiogenic effect equivalent to cell therapy. Methodology/Principal Findings: EPC-derived conditioned medium (EPC-CM) was obtained from culture expanded EPC subjected to 72 hours of hypoxia. In vitro, EPC-CM significantly inhibited apoptosis of mature endothelial cells and promoted angiogenesis in a rat aortic ring assay. The therapeutic potential of EPC-CM as compared to EPC transplantation was evaluated in a rat model of chronic hindlimb ischemia. Serial intramuscular injections of EPC-CM and EPC both significantly increased hindlimb blood flow assessed by laser Doppler (81.2 +/- 2.9% and 83.7 +/- 3.0% vs. 53.5 +/- 2.4% of normal, P < 0.01) and improved muscle performance. A significantly increased capillary density (1.62 +/- 0.03 and 1.68 +/- 0.05/muscle fiber, P < 0.05), enhanced vascular maturation (8.6 +/- 0.3 and 8.1 +/- 0.4/HPF, P < 0.05) and muscle viability corroborated the findings of improved hindlimb perfusion and muscle function. Furthermore, EPC-CM transplantation stimulated the mobilization of bone marrow (BM)-derived EPC compared to control (678.7 +/- 44.1 vs. 340.0 +/- 29.1 CD34(+)/CD45(-) cells/1x10(5) mononuclear cells, P < 0.05) and their recruitment to the ischemic muscles (5.9 +/- 0.7 vs. 2.6 +/- 0.4CD34(+) cells/HPF, P < 0.001) 3 days after the last injection. Conclusions/Significance: Intramuscular injection of EPC-CM is as effective as cell transplantation for promoting tissue revascularization and functional recovery. Owing to the technical and practical limitations of cell therapy, cell free conditioned media may represent a potent alternative for therapeutic angiogenesis in ischemic cardiovascular diseases.
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页数:11
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