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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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Submitted October 25, 2006
Accepted March 20, 2007
Identification of functional endothelial progenitor cells suitable for the treatment of ischemic tissue using human umbilical cord blood
Masumi Nagano, Toshiharu Yamashita, Hiromi Hamada, Kinuko Ohneda, Ken-ichi Kimura, Tomoki Nakagawa, Masabumi Shibuya, Hiroyuki Yoshikawa, and Osamu Ohneda*
Department of Regenerative Medicine, Graduate School of Comprehensive Sciences, University of Tsukuba, Tsukuba, Japan
Department of Obstetrics and Gynecology, Graduate School of Comprehensive Sciences, University of Tsukuba, Tsukuba, Japan
Laboratory of Molecular Pathophysiology, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan
Division of Genetics, Institute of Medical Science, University of Tokyo, Tokyo, Japan
* Corresponding author; email: oohneda{at}md.tsukuba.ac.jp.
Umbilical cord blood (UCB) has been utilized as a potential source of various kinds of stem cells, including hematopoietic stem cells, mesenchymal stem cells, and endothelial progenitor cells (EPC), for a variety of cell therapies. Recently, EPC were introduced for restoring vascularization in ischemic tissues. An appropriate procedure for isolating EPC 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 EPC from UCB by a combination of negative immunoselection and cell culture techniques. In addition, we divided EPC into two subpopulations according to the aldehyde dehydrogenase (ALDH) activity. We found that EPC 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 upregulated and VEGF, CXCR4, and Glut-1 mRNAs are increased in Alde-Low EPC under hypoxic conditions, while the response was not significant in Alde-High EPC. In fact, the introduction of Alde-Low EPC significantly reduced tissue damage in ischemia in a mouse flap model. Thus, the introduction of Alde-Low EPC may be a potential strategy for inducing rapid neovascularization and subsequent regeneration of ischemic tissues.
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