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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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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Figure 3

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Development of spindle-shaped and AT cells, and cord-like structures fro...
Development of spindle-shaped and AT cells, and cord-like structures from MNCCD34+. (a) When labeled MNCCD34– alone were plated on fibronectin, most of the cells did not attach to fibronectin and had a round shape. (b) When labeled MNCCD34+ alone were plated on fibronectin, only limited numbers of cell clusters and AT cells appeared within 7 days. (c) When green fluorescence-labeled MNCCD34– were coplated with unlabeled MNCCD34+, fluorescence-positive MNCCD34– participated in cell cluster formation, but they were located only at the center of the clusters. AT cells and cord-like structures were mostly fluorescence negative. (d) Conversely, when fluorescence-labeled MNCCD34+ were coplated with unlabeled MNCCD34–, the AT cells, cell clusters, and cord-like structures were mostly positive for the fluorescence. (e, f) From coculture of fluorescence-labeled MNCCD34+ and unlabeled MNCCD34–, numerous fluorescence-positive cell clusters developed, and some of the cell clusters connected together to form linear cord-like structures. Bars: 20 μm in a–d, 100 μm in e and f. (g) Bar graphs showing the numbers of AT cells and cell clusters developed from four different conditions of cell culture (1,000 cells/mm2). A limited number of AT cells and cell clusters appeared from MNCCD34+ alone. The numbers of developed AT cells and cell clusters were greatly enhanced by coculture of MNCCD34+ with MNCCD34– when compared with culture of MNCCD34+ alone. The numbers of AT cells and cell clusters derived from coculture of MNCCD34+ and MNCCD34– were comparable to those from total MNCs.