内皮克隆形成细胞旁分泌对人脐静脉内皮细胞生物学功能的影响
发布时间：2019-04-10 08:30
【摘要】：背景:研究表明,内皮克隆形成细胞移植对缺血损伤组织的血管新生具有促进作用。然而,这一作用是否与其旁分泌效应有关,仍有待进一步研究证实。目的:检测人脐血来源内皮克隆形成细胞条件培养基中细胞因子分泌谱,并观察内皮克隆形成细胞条件培养基对人脐静脉内皮细胞增殖、迁移及成管能力的影响。方法:从人脐血中分离、培养内皮克隆形成细胞,并进行细胞鉴定。采用抗体芯片对无血清内皮克隆形成细胞条件培养基中的细胞因子进行检测。采用无血清EBM-2培养基作为对照组,应用CCK-8、细胞划痕实验、Matrigel成管实验检测内皮克隆形成细胞条件培养基对人脐静脉内皮细胞增殖、迁移及成管能力的影响。结果与结论:(1)原代培养细胞形态:培养的细胞呈"铺路石"样生长,表达CD34,KDR及CD144,不表达CD45及CD133,Dil-acLDL摄取及FITC-UEA-I结合双阳性,在Matrigel基质胶中可形成管腔样结构,以上均符合内皮克隆形成细胞的特征;(2)抗体芯片检测:内皮克隆形成细胞条件培养基高表达30种促血管新生因子;(3)CCK-8检测:实验组人脐静脉内皮细胞在24,48,72 h增殖活性均高于对照组(P0.01);(4)细胞划痕实验结果显示,实验组人脐静脉内皮细胞迁移率在12,24 h均高于对照组(P0.01);(5)与对照组相比,实验组人脐静脉内皮细胞在Matrigel基质胶上可形成更多的管状结构(P0.01)。(6)结果表明:内皮克隆形成细胞能通过旁分泌效应促进成熟内皮细胞的生物活性。
[Abstract]:Background: studies have shown that endothelial clone-forming cell transplantation can promote angiogenesis in ischemic tissue. However, whether this effect is related to paracrine effect needs further study. Aim: to detect the cytokine secretion profile in the conditioned medium of human umbilical cord blood derived endothelial clone forming cells, and to observe the effects of endothelial clone forming cell conditioned medium on the proliferation, migration and tubular ability of human umbilical vein endothelial cells (HUVECs). Methods: the cells were isolated from human umbilical cord blood, cultured and identified. The cytokines in serum-free endothelial clone forming cell conditioned medium were detected by antibody microarray. Serum-free EBM-2 medium was used as control group, CCK-8, cell scratch test and Matrigel tube forming test were used to detect the effect of endothelial clone forming cell conditioned medium on the proliferation, migration and tube-forming ability of human umbilical vein endothelial cells (HUVECs). Results & conclusion: (1) Primary culture cell morphology: the cultured cells grew like "paving stone", the expression of CD34,KDR and CD144, did not express CD45 and CD133,Dil-acLDL uptake, and FITC-UEA-I binding double positive. A tube-like structure could be formed in the Matrigel matrix gel, all of which were consistent with the characteristics of endothelial clone-forming cells. (2) Detection of antibody chip: endothelial clone formed cell culture medium with high expression of 30 proangiogenic factors; (3) CCK-8 assay: the proliferation activity of human umbilical vein endothelial cells in the experimental group was higher than that in the control group at 24,48,72 h (P0.01); (4) the results of cell scratch test showed that the migration rate of human umbilical vein endothelial cells in the experimental group was higher than that in the control group at 12 h and 24 h (P0.01). (5) compared with the control group, Experimental group of human umbilical vein endothelial cells could form more tubular structure on Matrigel matrix gel (P0.01). (6). The results showed that endothelial clone forming cells could promote the biological activity of mature endothelial cells through paracrine effect.
【作者单位】： 南方医科大学;解放军广州军区广州总医院;广东省中山市第二人民医院;
【基金】：国家自然科学基金面上项目(81571910) 广东省科技计划项目(2014A020212256) 广州市科技计划项目(201607010394)~~
【分类号】：R329.2
本文编号：2455647
[Abstract]:Background: studies have shown that endothelial clone-forming cell transplantation can promote angiogenesis in ischemic tissue. However, whether this effect is related to paracrine effect needs further study. Aim: to detect the cytokine secretion profile in the conditioned medium of human umbilical cord blood derived endothelial clone forming cells, and to observe the effects of endothelial clone forming cell conditioned medium on the proliferation, migration and tubular ability of human umbilical vein endothelial cells (HUVECs). Methods: the cells were isolated from human umbilical cord blood, cultured and identified. The cytokines in serum-free endothelial clone forming cell conditioned medium were detected by antibody microarray. Serum-free EBM-2 medium was used as control group, CCK-8, cell scratch test and Matrigel tube forming test were used to detect the effect of endothelial clone forming cell conditioned medium on the proliferation, migration and tube-forming ability of human umbilical vein endothelial cells (HUVECs). Results & conclusion: (1) Primary culture cell morphology: the cultured cells grew like "paving stone", the expression of CD34,KDR and CD144, did not express CD45 and CD133,Dil-acLDL uptake, and FITC-UEA-I binding double positive. A tube-like structure could be formed in the Matrigel matrix gel, all of which were consistent with the characteristics of endothelial clone-forming cells. (2) Detection of antibody chip: endothelial clone formed cell culture medium with high expression of 30 proangiogenic factors; (3) CCK-8 assay: the proliferation activity of human umbilical vein endothelial cells in the experimental group was higher than that in the control group at 24,48,72 h (P0.01); (4) the results of cell scratch test showed that the migration rate of human umbilical vein endothelial cells in the experimental group was higher than that in the control group at 12 h and 24 h (P0.01). (5) compared with the control group, Experimental group of human umbilical vein endothelial cells could form more tubular structure on Matrigel matrix gel (P0.01). (6). The results showed that endothelial clone forming cells could promote the biological activity of mature endothelial cells through paracrine effect.
【作者单位】： 南方医科大学;解放军广州军区广州总医院;广东省中山市第二人民医院;
【基金】：国家自然科学基金面上项目(81571910) 广东省科技计划项目(2014A020212256) 广州市科技计划项目(201607010394)~~
【分类号】：R329.2
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