从脐血单个核细胞诱导巨核细胞体外扩增及分化的实验研究

发布时间：2018-01-15 22:11

  本文关键词：从脐血单个核细胞诱导巨核细胞体外扩增及分化的实验研究　出处：《福建医科大学》2005年硕士论文　论文类型：学位论文

  更多相关文章： 脐血 巨核细胞 细胞因子 体外扩增 诱导分化

【摘要】：巨核细胞是血小板的前体细胞。将部分脐血进行巨核系祖细胞体外扩增后再输注给患者,是有望解决脐血造血干细胞移植后患者血小板恢复迟缓这一难题的有效途径,但目前最佳扩增条件的确立仍处于实验阶段。本项研究从脐血单个核(MNC)出发,在无血清培养体系中比较了TPO、TPO+SCF、TPO+SCGF、TPO+IL-3、TPO+EPO5 种简单细胞因子组合对刺激巨核细胞扩增的作用,经体外培养14 天后,TPO+SCF 组CD41~+细胞比例和扩增倍数最高,分别为20.95±2.83%和48.40±5.71倍;接着,将TPO 组和TPO+SCF 组的脐血MNC 均培养到18 天,以观察培养后期(9d、14d、18d)巨核细胞的凋亡情况,结果发现TPO+SCF 组的MNC 和CD41~+细胞的凋亡率与单用TPO 组相比均有显著差异(P0.05);最后,以TPO+SCF组为研究对象,采用RT-PCR 方法检测了转录因子GATA-1、NF-E2 和系特异抗原GPⅡb mRNA 表达情况,发现培养14 天时三者的表达均达到高峰。上述结果证明:SCF 和TPO 的协同刺激在有效促进巨核细胞的扩增的同时,也在有效诱导其分化。
[Abstract]:Megakaryocytes are progenitor cells of platelets. Some umbilical cord blood was amplified by megakaryocyte progenitor cells in vitro and then infused to patients. It is an effective way to solve the problem of delayed platelet recovery after cord blood hematopoietic stem cell transplantation. However, the optimal amplification conditions are still in the experimental stage. This study compared TPO SCF in serum-free culture system from cord blood mononuclear cells (MNCs). The effect of simple cytokine combination of TPO SCGF- TPO IL-3 TPO EPO5 on the proliferation of megakaryocytes cultured in vitro for 14 days. CD41 ~ ~ cells in TPO SCF group were 20.95 卤2.83% and 48.40 卤5.71 times respectively. Then, the cord blood MNC of TPO group and TPO SCF group were cultured for 18 days to observe the apoptosis of megakaryocyte in the late stage of culture. The results showed that the apoptosis rates of MNC and CD41 ~ ~ cells in TPO SCF group were significantly different from those in TPO group. Finally, the expression of transcription factor GATA-1NF-E2 and lineage-specific antigen GP 鈪，

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