摘要
本研究探讨血小板衍生微粒(PMP)对脐血粒巨噬祖细胞集落形成单位(CFU-GM)的促增殖作用。采用不同浓度的凝血酶激活血小板释放出PMP,采用流式细胞术检测PMP的释放量,确定凝血酶的最佳实验浓度。应用Ficoll分离脐血单个核细胞(MNC),在含有不同浓度PMP的甲基纤微素半固体培养基中培养7天,观察CFU-GM集落形成情况。结果表明:2.0、1.5、1.0和0.5U/ml凝血酶激活血小板后的PMP释放率分别为:28.7%、47.7%、50.1%和43.9%;CFU-GM集落产率与PMP呈剂量依赖关系,PMP为10、50、100μg/ml时的CFU-GM集落产率(个/2×105MNC)分别为:119.8±32.2、142.8±45.2、180.8±85.1。50、100μg/mlPMP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/mlPMP组的集落产率略高于空白对照组,但P值>0.05。结论:用1.0U/ml的凝血酶激活血小板释放的PMP较为均一,而且释放量最大。PMP可促进人脐血粒巨噬祖细胞的增殖。
This study was aimed to investigate the effect of platelet-derived microparticles （PMP） on stimulating the proliferation of granulocyte-macrophage progenitors （CFU-GM） from umbilical cord blood. Different concentrations of thrombin were adopted to activate the platelets for releasing PMP. Flow cytometry was adopted to evaluate the efficiencies of different concentrations of thrombin to produce PMP. Umbilical cord blood mononuclear cells （MNC） were obtained from healthy donors and the MNC were isolated by Ficoll density gradient centrifugation. MNC were cultured in 2.7% methylcellulose containing different concentration of PMP and colonies were counted under an inverted microscope after 7 days. The result showed that the release rate of PMP activated by 2.0,1.5,1.0 and 0.5 U/ml thrombin were 28. 7%, 47.7%, 50.1% and 43.9% respectively. The PMP enhanced colony formation in dose-dependent manner. The number of colonies per 2×10^5 MNCs in groups of PMP at different concentrations （ 10, 50 and 100μg/ml） were 119.8±32.2,142.8±45.2 and 180.8±85.1 respectively. The number of colonies in the groups of PMP at 100μg/ml and 50μg/ml were statistically significant when compared with control group （ 103.0±24.8 ） （ P 〈 0.05 ）. The number of colonies per 2×10^5 MNC in the group of PMP （ 10μg/ml） was 119.8±32.2 which was higher than that in control group, but there was no statistical significance between two groups. It is concluded that platelet activated with 1.0 U/ml thrombin can get the best release efficiency of PMP and PMP can enhance the proliferation of granulocyte-macrophage progenitor cells of umbilical cord blood.
出处
《中国实验血液学杂志》
CAS
CSCD
2006年第3期561-564,共4页
Journal of Experimental Hematology
基金
江苏省卫生厅科研课题
编号:H200533
关键词
血小板衍生微粒
脐血
粒巨噬祖细胞
platelet-derived microparticles
cord blood
granulocyte-macrophage progenitor cell
