Peripheral blood progenitor cells to improve the study of limb ischemia.docVIP

 PAGE \* MERGEFORMAT 2 Peripheral blood progenitor cells to improve the study of limb ischemia Author: Wang Hui, Wang Ling, Li Kai-Zong, Rui Ling, Sun Baohua, Li Xiaojun, Yi-jun [Keywords:] endothelium; stem cells; transplantation; peripheral blood [Abstract] AIM: To estimate the effect of human peripheral blood derivedendothelial progenitor cell (EPC) transplantation on the repairment of limb ischemia. METHODS: The mononuclear cells from human peripheral blood were cultured in M199 medium supplemented with 10% fetal bovine serum, 20 ng / mL VEGF and 4ng/mL bFGF in vitro. The specific markers of EPC were analyzed by FCS at day 6 and 14 during culture respectively. The bilateral femoral artery, great saphenous artery, iliac circumflex artery, and muscular branch of 15 Balb / c nude mice were ligated to cause limb ischemia. The 6day adherence cells marked by CMDiI were multipoint injected into the left local muscles of ischemic limbs (EPC group) and M199 was multipoint injected into the right local muscles of ischemic limbs (M199 group) in order to evaluate the therapeutic effect. Seven days after operation, 60 μ g FITCUEA was injected via tail vein.One hour later, the mice were killed.The heart, liver, lung, spleen, kidney and limb gastrocnemius were taken and examined with fluorescence microscopy . The number of capillaries of ischemic limbs was measured by factor Ⅷ related antigen immunohistochemical staining. RESULTS: The cultured cells at day 6 expressed KDR, CD133, CD34 and vWF, and the positive rates of KDR, CD133, CD34 and vWF were (46.4 ± 9.3)%, (33.8 ± 11.7)%, (73.5 ± 6.3)% and (38.9 ± 8.2)%, respectively. At the 14th day during culture, the cells expressed KDR, CD34 and vWF and the positive rates of KDR, CD34 and vWF were (81.5 ± 7.6)%, (88.9 ± 5.4)% and (78.3 ± 13.6)%, respectively; the positive rate of CD133 dropped to (3.8 ± 8.7)%. The necrosis and density of capillary of the ischemic limbs of nude mice were improved more obviously in