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 Blood, 1 July 2007, Vol. 110, No. 1, pp. 151-160.
Prepublished online as a Blood First Edition Paper on March 22, 2007; DOI 10.1182/blood-2006-10-047092.

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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY

Identification of functional endothelial progenitor cells suitable for the treatment of ischemic tissue using human umbilical cord blood

Masumi Nagano1, Toshiharu Yamashita1, Hiromi Hamada2, Kinuko Ohneda3, Ken-ichi Kimura1, Tomoki Nakagawa1, Masabumi Shibuya4, Hiroyuki Yoshikawa2, and Osamu Ohneda1

Graduate School of Comprehensive Human Sciences1 Department of Regenerative Medicine, 2 Department of Obstetrics and Gynecology, University of Tsukuba, Japan; 3 Laboratory of Molecular Pathophysiology, Faculty of Pharmacy, Takasaki University of Health and Welfare, Japan; 4 Division of Genetics, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo, Japan

Umbilical cord blood (UCB) has been used as a potential source of various kinds of stem cells, including hematopoietic stem cells, mesenchymal stem cells, and endothelial progenitor cells (EPCs), for a variety of cell therapies. Recently, EPCs were introduced for restoring vascularization in ischemic tissues. An appropriate procedure for isolating EPCs from UCB is a key issue for improving therapeutic efficacy and eliminating the unexpected expansion of nonessential cells. Here we report a novel method for isolating EPCs from UCB by a combination of negative immunoselection and cell culture techniques. In addition, we divided EPCs into 2 subpopulations according to the aldehyde dehydrogenase (ALDH) activity. We found that EPCs with low ALDH activity (Alde-Low) possess a greater ability to proliferate and migrate compared to those with high ALDH activity (Alde-High). Moreover, hypoxia-inducible factor proteins are up-regulated and VEGF, CXCR4, and GLUT-1 mRNAs are increased in Alde-Low EPCs under hypoxic conditions, while the response was not significant in Alde-High EPCs. In fact, the introduction of Alde-Low EPCs significantly reduced tissue damage in ischemia in a mouse flap model. Thus, the introduction of Alde-Low EPCs may be a potential strategy for inducing rapid neovascularization and subsequent regeneration of ischemic tissues.


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