The adipose-derived stromal vascular fraction (SVF) is an effective source for autologous cell transplantation. However, the quality and quantity of SVFs vary depending on the patient’s age, complications, and other factors. In this study, we developed a method to reproducibly increase the cell number and improve the quality of adipose-derived SVFs by surgical procedures, which we term “wound repair priming.” Subcutaneous fat from the inguinal region of BALB/c mice was surgically processed (primed) by mincing adipose parenchyma (injury) and ligating the subcutaneous fat-feeding artery (ischemia). SVFs were isolated on day 0, 1, 3, 5, or 7 after the priming procedures. Gene expression levels of the primed SVFs were measured via microarray and pathway analyses which were performed for differentially expressed genes. Changes in cellular compositions of primed SVFs were analyzed by flow cytometry. SVFs were transplanted into syngeneic ischemic hindlimbs to measure their angiogenic and regeneration potential. Hindlimb blood flow was measured using a laser Doppler blood perfusion imager, and capillary density was quantified by CD31 staining of ischemic tissues. Stabilization of HIF-1 alpha and VEGF-A synthesis in the SVFs were measured by fluorescent immunostaining and Western blotting, respectively. As a result, the number of SVFs per fat weight was increased significantly on day 7 after priming. Among the differentially expressed genes were innate immunity-related signals on both days 1 and 3 after priming. In primed SVFs, the CD45-positive blood mononuclear cell fraction decreased, and the CD31-CD45-double negative mesenchymal cell fraction increased on day 7. The F4/80-positive macrophage fraction was increased on days 1 and 7 after priming. There was a serial decrease in the mesenchymal-gated CD34-positive adipose progenitor fraction and mesenchymal-gated CD140A-positive/CD9-positive preadipocyte fraction on days 1 and 3. Transplantation of primed SVFs resulted in increased capillary density and augmented blood flow, improving regeneration of the ischemic limbs. HIF-1 alpha was stabilized in the primed cutaneous fat in situ, and VEGF-A synthesis of the primed SVFs was on a peak on 5 days after priming. Wound repair priming thus resulted in SVFs with increased number and augmented angiogenic potential.

中文翻译：

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在小鼠肢体缺血模型中，外科手术损伤和局部缺血可引发脂肪基质血管部分并增加血管生成能力。

脂肪来源的基质血管部分（SVF）是自体细胞移植的有效来源。但是，SVF的质量和数量取决于患者的年龄，并发症和其他因素。在这项研究中，我们开发了一种方法，该方法通过外科手术程序可复制地增加细胞数量并改善脂肪来源的SVF的质量，我们将其称为“伤口修复启动”。通过切碎脂肪实质（损伤）并结扎皮下供脂动脉（局部缺血），对BALB / c小鼠腹股沟区的皮下脂肪进行手术处理（预涂）。在启动程序后第0、1、3、5或7天分离出SVF。通过微阵列和途径分析测量了引发的SVF的基因表达水平，对差异表达的基因进行了分析。通过流式细胞仪分析引发的SVF的细胞组成的变化。SVFs被移植到同基因缺血性后肢中，以测量其血管生成和再生潜力。使用激光多普勒血流灌注成像仪测量后肢血流量，并通过缺血组织的CD31染色定量毛细管密度。分别通过荧光免疫染色和蛋白质印迹法测量了SVF中HIF-1α和VEGF-A合成的稳定性。结果，在启动后第7天，每脂肪重量的SVF数量显着增加。在差异表达的基因中，有在引发后第1天和第3天都有与先天性免疫相关的信号。在引发的SVF中，第7天的CD45阳性血液单核细胞分数降低，而CD31-CD45-双阴性间充质细胞分数增加。启动后第1天和第7天F4 / 80阳性巨噬细胞分数增加。在第1天和第3天，间质门控的CD34阳性脂肪祖细胞部分和间质门控的CD140A阳性/ CD9阳性前脂肪细胞部分减少了。缺血肢体的再生。HIF-1α在引发的皮肤脂肪中稳定 改善缺血肢体的再生。HIF-1α在引发的皮肤脂肪中稳定 改善缺血肢体的再生。HIF-1α在引发的皮肤脂肪中稳定原位，涂底漆的SVFs和VEGF-A的合成引发后是在峰值上5天。因此，伤口修复引发的SVF数量增加，血管生成潜力增强。