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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 to investigate the effect of platelet-derived microparticles (PMP) on the proliferation of cord blood monocyte-derived macrophage progenitor cells (CFU-GM). The platelets were activated by different concentrations of thrombin to release PMP. The release of PMP was determined by flow cytometry and the optimal concentration of thrombin was determined. Umbilical cord blood mononuclear cells (MNCs) were isolated by Ficoll and cultured in methylcellulose semi-solid medium containing different concentrations of PMP for 7 days to observe the CFU-GM colony formation. The results showed that the release rates of PMP after platelets activated by thrombin 2.0,1.5,1.0 and 0.5U / ml were 28.7%, 47.7%, 50.1% and 43.9%, respectively. The CFU-GM production rate was in a dose-dependent manner with PMP, The CFU-GM colony yields (one / 2 × 10 5 MNC) at PMP 10, 10 and 100 μg / ml were 119.8 ± 32.2, 142.8 ± 45.2, 180.8 ± 85.1.50 and 100 μg / ml, respectively. P value was less than 0.05 in blank control group (103.0 ± 24.8), while that in 10 μg / ml PMP group was slightly higher than that in blank control group (P> 0.05). CONCLUSION: PMP released from platelets activated with 1.0 U / ml thrombin is more uniform and has the highest release. PMP can promote the proliferation of human umbilical cord blood macrophage progenitor cells.