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1 Division of Cardiovascular Surgery and Medicine, Department of Medical Bioregulation, Yamaguchi University School of Medicine, Ube, Yamaguchi, Japan
* To whom correspondence should be addressed. E-mail: kimikazu{at}yamaguchi-u.ac.jp.
Ex vivo expansion of stem cells might be a feasible method of resolving the problem of limited cell supply in cell-based therapy. The implantation of expanded CD34+ endothelial progenitor cells has the capacity to induce angiogenesis. In this study, we tried to induce angiogenesis by implanting expanded CD117+ stem cells derived from mouse bone marrow.
After 2 weeks of culture with the addition of several growth factors, the CD117+ stem cells
expanded to about 20-fold, and had endothelial phenotype with high expression of CD34 and VE-cadherin. However, more than 70% of these ex vivo expanded cells had a senescent
phenotype by 
-galactosidase staining, and their survival and incorporation were poor after implantation into the ischemic limbs of mice. Compared with the PBS injection only, the microvessel density and the percentage of limb blood flow were significantly higher after the
implantation of 2 x 105 freshly collected CD117+ cells (P < 0.01), but not after the
implantation of 2 x 105 expanded CD117+ cells (P > 0.05). These data indicate that ex vivo expansion of CD117+ stem cells has low potency for inducing therapeutic angiogenesis, which might be related to the cellular senescence during ex vivo expansion.
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