地西他滨对激素抵抗型ITP患者骨髓巨核细胞培养的影响

Effect of Decitabine on Megakaryocyte Culture of Steroid-resistant ITP Patients

目的:探讨地西他滨和ITP患者血浆对激素抵抗型ITP患者骨髓巨核细胞培养的影响。方法:收集20例激素抵抗型ITP患者骨髓并分离出单个核细胞,采用甲基纤维素半固体培养法(观察并计数巨核细胞集落)和液体培养法(检测CD41a+细胞表达率)进行巨核细胞培养,根据培养条件的不同将实验分成4组:A组为空白对照组,B组为地西他滨组,C组为ITP患者血浆组,D组为地西他滨+ITP患者血浆组。除上述成分不同外,4组中其余培养条件均相同。在倒置显微镜下观察并计数巨核细胞集落,用流式细胞术检测所培养细胞中CD41a+细胞的表达率。结果:不同浓度的地西他滨对巨核细胞集落生长的作用不同,地西他滨浓度为3.0μmol/L时能明显促进巨核细胞生长。B组与A组相比,巨核细胞集落数明显增加,CD41a+细胞表达率也较高,差异有统计学意义(P<0.05)。C组与A组相比,巨核细胞集落数明显降低,差异有统计学意义。D组与C组相比,巨核细胞集落数较多,其差异有统计学意义。结论:地西他滨在一定浓度范围内能促进激素抵抗型ITP患者骨髓巨核细胞集落生长,且以浓度为3.0μmol/L时,巨核细胞集落最多。ITP患者血浆对激素抵抗型ITP患者骨髓巨核细胞生长有抑制作用。

Objective : To investigate the effect of decitabine and plasma of ITP patients on in vitro cultrue of megakaryocytes in bone marrowof steroid-resistant ITP patients. Me thods: Bone marrowmononuclear cells were isolated from 20 steroid-resistant ITP patients,both methyl cellulose semisolid culture system( to observe and count the number of megakaryocytes colony-forming unit) and liquid culture system( to analysis the expression rate of CD41a+cells) were used for megakaryocyte cultrue. The experiments were divided into 4 groups according to the different components of the culture system,group A was control,group B was added with decitabine,group C with ITP plasma,group D with both decitabine and ITP plasma,and the rest of the culture components were the same in the 4 groups except the above-mentioned materials. M orphology of megakaryocytes was observed by inverted and light microscopy.The expression rate of CD41a+cells in culture was analysed by flowcytometric. Re sults: Different concentration of decitabine showed different effect on megakaryocyte growth of steroid-resistant ITP patients and the optimal concentration to differentiate into megakaryocyte for bone marrowmononuclear cells is 3. 0 μmol / L. Compared with group A,both megakaryocyte colony forming units( CFU) and expression rate of CD41a+cells in group B were statistically significantly higher( P < 0. 05). As compared with group A,the megakaryocyte colony-forming units in group C decreased with statistically significant difference,while compared with group C,the megakaryocyte colony-forming units in group D obviously increased with statistically significant difference. Conclusions: Decitabine is able to induce bone marrowmononuclear cells of steroid-resistant ITP patients to differentiate into megakaryocyte and the optimal concentration is 3. 0 μmol / L; ITP plasma is able to inhibit the megakaryocyte growth of steroid-resistant ITP patients.