Transplanted cord blood–derived endothelial precursor cells augment postnatal neovascularization
Toyoaki Murohara, … , Kazuo Matsui, Tsutomu Imaizumi
Toyoaki Murohara, … , Kazuo Matsui, Tsutomu Imaizumi
Published June 1, 2000
Citation Information: J Clin Invest. 2000;105(11):1527-1536. https://doi.org/10.1172/JCI8296.
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Article

Transplanted cord blood–derived endothelial precursor cells augment postnatal neovascularization

Abstract

Endothelial precursor cells (EPCs) have been identified in adult peripheral blood. We examined whether EPCs could be isolated from umbilical cord blood, a rich source for hematopoietic progenitors, and whether in vivo transplantation of EPCs could modulate postnatal neovascularization. Numerous cell clusters, spindle-shaped and attaching (AT) cells, and cord-like structures developed from culture of cord blood mononuclear cells (MNCs). Fluorescence-trace experiments revealed that cell clusters, AT cells, and cord-like structures predominantly were derived from CD34-positive MNCs (MNCCD34+). AT cells and cell clusters could be generated more efficiently from cord blood MNCs than from adult peripheral blood MNCs. AT cells incorporated acetylated-LDL, released nitric oxide, and expressed KDR, VE-cadherin, CD31, and von Willebrand factor but not CD45. Locally transplanted AT cells survived and participated in capillary networks in the ischemic tissues of immunodeficient nude rats in vivo. AT cells thus had multiple endothelial phenotypes and were defined as a major population of EPCs. Furthermore, laser Doppler and immunohistochemical analyses revealed that EPC transplantation quantitatively augmented neovascularization and blood flow in the ischemic hindlimb. In conclusion, umbilical cord blood is a valuable source of EPCs, and transplantation of cord blood–derived EPCs represents a promising strategy for modulating postnatal neovascularization.

Authors

Toyoaki Murohara, Hisao Ikeda, Junli Duan, Satoshi Shintani, Ken-ichiro Sasaki, Hiroyuki Eguchi, Ichiro Onitsuka, Kazuo Matsui, Tsutomu Imaizumi

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Augmented neovascularization by local transplantation of either umbilica...
Augmented neovascularization by local transplantation of either umbilical cord blood–derived (UCB-derived) or adult peripheral blood–derived (APB-derived) EPCs in the ischemic hindlimb of immunodeficient nude rats. (a) Unilateral (left) hindlimb ischemia (arrowheads) was surgically induced in nude rats. Either APB-derived EPCs (3 × 105 cells/animal, n = 6), UCB-derived EPCs (3 × 105 cells/animal, n = 8), or saline (control, n = 7) were locally transplanted into the ischemic hindlimb at postoperative day 3. Representative serial LDPI analyses showed progressive increases in blood flow in the ischemic hindlimb (red to white color) of rats injected with EPCs obtained from APB or UCB. In contrast, recovery of blood flow was retarded in the ischemic limb (blue to green color) in a control animal. (b) Computer-assisted quantitative analyses of hindlimb blood flow demonstrated significantly enhanced ischemic/normal limb blood perfusion ratios in rats injected with EPCs of either origin (APB or UCB) compared with controls. AP < 0.001. (c) Capillary ECs were identified in the sections of ischemic tissues under a light microscope after immunohistochemical staining for vWF (brown reaction products) and alkaline phosphatase (AP) staining (blue reaction products). Light microscopy revealed increased capillary densities in rats receiving EPCs of either origin (APB or UCB) compared with controls. Bars, 50 μm. (d) Quantitative analyses showed increased capillary densities in the ischemic hindlimb tissues of the two EPC-transplanted groups as compared with control animals examined 14 days after induction of limb ischemia. AP < 0.001.