Liao You-Ping, Liu Yu-Xia, Li Qing-Zhao, Hu Guo-Yu
Department of Hematology, Zhuzhou Central Hospital, Zhuzhou 412007, Hunan Province, China.
Department of Hematology, Zhuzhou Central Hospital, Zhuzhou 412007, Hunan Province, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Aug;23(4):1092-6. doi: 10.7534/j.issn.1009-2137.2015.04.036.
To investigate the effect of decitabine and plasma of ITP patients on in vitro cultrue of megakaryocytes in bone marrow of steroid-resistant ITP patients.
Bone marrow mononuclear 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. Morphology of megakaryocytes was observed by inverted and light microscopy. The expression rate of CD41a⁺ cells in culture was analysed by flow cytometric.
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 marrow mononuclear 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.
Decitabine is able to induce bone marrow mononuclear 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.
探讨地西他滨及免疫性血小板减少症（ITP）患者血浆对激素抵抗型ITP患者骨髓巨核细胞体外培养的影响。
从20例激素抵抗型ITP患者中分离出骨髓单个核细胞，采用甲基纤维素半固体培养体系（观察并计数巨核细胞集落形成单位数量）和液体培养体系（分析CD41a(+)细胞的表达率）进行巨核细胞培养。根据培养体系不同成分将实验分为4组，A组为对照组，B组添加地西他滨，C组添加ITP患者血浆，D组同时添加地西他滨和ITP患者血浆，除上述物质外，4组其余培养成分相同。通过倒置显微镜和光学显微镜观察巨核细胞形态。采用流式细胞术分析培养物中CD41a⁺细胞的表达率。
不同浓度的地西他滨对激素抵抗型ITP患者巨核细胞生长的影响不同，骨髓单个核细胞分化为巨核细胞的最佳浓度为3.0 μmol/L。与A组相比，B组巨核细胞集落形成单位（CFU）和CD41a⁺细胞表达率均有统计学显著升高（P < 0.05）。与A组相比，C组巨核细胞集落形成单位减少，差异有统计学意义；与C组相比，D组巨核细胞集落形成单位明显增加，差异有统计学意义。
地西他滨能够诱导激素抵抗型ITP患者的骨髓单个核细胞分化为巨核细胞，最佳浓度为3.0 μmol/L；ITP患者血浆能够抑制激素抵抗型ITP患者巨核细胞的生长。
