外周血内皮祖细胞移植对肢体缺血的改善作用

背景:内皮祖细胞移植为肢体缺血的治疗提供了新的选择。目的:研究人外周血来源内皮祖细胞移植对改善肢体缺血的作用。方法:采用Ficoll密度梯度离心法获取人外周血单个核细胞,体外诱导扩增6d后,检测其内皮祖细胞特异性标志的表达,并将荧光染料标记后的贴壁细胞通过缺血局部多点注射移植到后肢缺血的裸鼠动物模型体内,以评价其治疗效果。结果与结论:从人外周血单个核细胞诱导出的贴壁细胞可表达内皮祖细胞特异性标志物CD133、CD34和血管内皮生长因子受体2,说明从人外周血单个核细胞中可诱导出内皮祖细胞。移植内皮祖细胞后裸鼠缺血后肢的坏死情况和毛细血管密度均明显改善(P<0.05);在缺血后肢肌肉石蜡切片中可见分散不均的红色和黄绿色荧光标记的内皮祖细胞的掺入。表明移植的内皮祖细胞可以定向整合到缺血局部,改善裸鼠的后肢缺血。

Transplantation of endothelial progenitor cells from human peripheral blood improves limb ischemia

BACKGROUND:Transplantation of endothelial progenitor cells provides us a new treatment options on limbs ischemia which is hard to cure in clinical.OBJECTIVE:To estimate the effect of transplantation of endothelial progenitor cells(EPCs) derived from human peripheral blood on the improvement of limbs ischemia.METHODS:The mononuclear cells derived from human peripheral blood were obtained by Ficoll density gradient centrifugation.The expressions of specific markers of EPCs were detected after cultured and amplified in vitro for 6 days,and the adherence cells marked by fluorochrome were multipoint injected into the nude mouse model with hind limb ischemia in order to evaluate the therapeutic effect.RESULTS AND CONCLUSION:The adherence cells derived from mononuclear cells could express CD133,CD34 and vascular endothelial growth factor 2(VEGFR-2).It indicated that we could culture and obtain the EPCs from mononuclear cells of human peripheral blood in vitro under certain condition.The necrosis and capillary density of the ischemia limbs of nude mice were improved obviously after EPCs transplantation(P < 0.05).Red and flavo-green fluorescently-labeled EPCs were observed in the paraffin section of ischemia limbs muscle.The transplanted EPCs can incorporate into the local capillary networks and improve the ischemia of posterior limb of nude mice.