血小板衍生膜微粒促人脐带血造血干细胞增殖的实验研究

发布时间：2018-02-24 14:19

本文关键词： 血小板衍生膜微粒脐带血粒-巨噬祖细胞集落增殖　出处：《东南大学》2006年硕士论文　论文类型：学位论文

【摘要】： 目的研究血小板衍生膜微粒(PMPs)的制备方法,并探讨PMPs对人脐带血造血干细胞增殖能力的影响。方法分别采用不同浓度的凝血酶激活血小板释放出PMPs,采用流式细胞仪检测PMPs的释放量,确定凝血酶最佳实验浓度。应用扫描电镜观察凝血酶激活血小板后PMPs的释放情况。采用Ficoll分离人脐带血单个核细胞(MNCs),在含有不同浓度PMPs或血小板的甲基纤维素半固体培养基中培养7天,观察人脐带血粒-巨噬祖细胞集落(CFU-GM)形成情况,并用流式细胞仪检测在含PMPs的无血清培养体系中培养7天后人脐带血MNCs的CD34抗原表达的情况。结结果用2.0 U/ml、1.5 U/ml、1.0 U/ml和0.5 U/ml凝血酶激活血小板后的PMPs释放率分别为:28.7%、47.7%、50.1%和43.9%;CFU-GM产率与PMPs呈剂量依赖关系,PMPs为10μg/ml、50μg/ml、100μg/ml和血小板为200×109/L时的CFU-GM产率(个/2×105 MNCs)分别为:119.8±32.2、142.9±45.2、180.8±85.1和136.5±43.6, 50μg/ml、100μg/ml PMPs和200×109/L血小板组的集落产率与空白对照组(103.0±24.8)相比,P值均0.05;而10μg/ml PMPs组的集落产率略高于空白对照组,但两者无统计学差异,P值0.05。无血清培养7天后,50μg/ml PMPs组CD34抗原表达率为:0.73%±0.74%,与空白对照组(0.43%±0.54%)相比,P值0.05。结论用1.0 U/ml的凝血酶激活血小板释放的PMPs较为均一且释放量最大。PMPs可促进人脐带血粒-巨噬祖细胞的增殖和CD34抗原的表达。
[Abstract]:Objective to study the preparation of platelet-derived membrane (PMPs) and the effect of PMPs on the proliferation of human umbilical cord blood hematopoietic stem cells. Methods different concentrations of thrombin were used to activate platelet to release PMPs. Flow cytometry was used to detect the release of PMPs. The optimal concentration of thrombin was determined. The release of PMPs after thrombin activation was observed by scanning electron microscope. Ficoll was used to isolate mononuclear cells from human umbilical cord blood and methylcellulose hemicellulose containing different concentrations of PMPs or platelets was obtained. Culture in solid medium for 7 days, The formation of human umbilical cord blood granulocyte-macrophage colony (CFU-GM) was observed, and the expression of CD34 antigen of MNCs was detected by flow cytometry in serum-free culture system containing PMPs for 7 days. The PMPs release rates were 47.70.1% and 43.9% in a dose-dependent relationship with 10 渭 g / ml 50 渭 g / ml 50 渭 g / ml 50 渭 g / ml CFU-GM and 200 脳 10 ~ (5) MNCs, respectively. The PMPs release rates were: 119.8 卤32.2142.9 卤45.2180.8 卤85.1 and 136.5 卤43.6ml, 50 渭 g / ml, 100 渭 g / ml PMPs and 50 渭 g / ml / ml, respectively119.8 卤32.2142.9 卤45.2180.8 卤85.1 and 136.5 卤43.6ml / ml, 50 渭 g / ml, 100 渭 g / ml PMPs and 50 渭 g / ml / ml / ml of platelets / platelets, 200 脳 109Mncml / ml and 200 脳 109Mncml / ml of platelets, respectively519.8 卤32.2142.9 卤45.2142.9 卤45.2180.8 卤85.1 and 136.5 卤43.60.50 渭 g / ml respectively. Compared with the control group (103.0 卤24.8), the colony production rate of the 200 脳 10 9 / L platelet group was 0.05, while the colony yield rate of the 10 渭 g / ml PMPs group was slightly higher than that of the blank control group, and that of the 10 渭 g / ml PMPs group was slightly higher than that of the blank control group. After 7 days of serum-free culture, the expression rate of CD34 antigen in 50 渭 g / ml PMPs group was 0.73% 卤0.74%, which was higher than that in the blank control group (0.43% 卤0.54%). Conclusion PMPs released from platelets activated by 1.0 U / ml thrombin can promote the proliferation of human umbilical cord blood granulocyte-macrophage and the expression of CD34 antigen.
【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

【参考文献】