不同细胞因子组合对脐血巨核细胞的体外扩增效应

Effect of different cytokine groups on expansion in vitro of megakaryocyte from cord blood

目的 探讨在体外液体培养条件下不同细胞因子组合促进脐血单个核细胞向巨核系增殖分化的效应。方法 将TPO(thrombopoietin)、SCF(stemcellfactor)、SCGF(stemcellgrowthfactor)、IL- 3、EPO(erythropoietin)等细胞因子组合成不同的实验组,在无血清液体培养体系中,体外扩增脐血单个核细胞 (CBMC)14d。扩增后于不同时间点用流式细胞仪检测CD61+细胞的比例,同时计数活细胞数,从而求得较好的细胞因子组合组。结果 经 14d培养后,SCF+TPO组扩增效果最好,活细胞总数和CD61+细胞数量分别增加了 3. 14±0. 62倍和 48. 40±5. 71倍,显著高于单用TPO组的 1. 81±0. 36倍和 18. 72±3. 73倍。TPO+IL 3组使CD61+细胞数量增加了 25. 82±3. 88倍,但扩增后CD61+细胞比例仅为 9. 34% ±1. 92%,低于TPO组的12. 21%±2. 27%;TPO+SCGF组和TPO+EPO组的扩增效果与单用TPO组相比无统计学差异 (P>0. 05)。结论 SCF能协同TPO增强巨核系造血,增加培养体系中巨核细胞的纯度和数量。

Objective To investigate the capacity of different cytokines combination in inducing cord blood mononuclear cells (CBMNC) to proliferate and differentiate in vitro committedly into megakaryocytes in liquid culture system.Methods CBMNC in different experimental groups in which TPO,SCF,SCGF,IL-3 and EPO were combined in various manner were expanded in a serum-free culture system for 14 days.The proportion of CBMNC with CD61 antigen expression was measured by FACS and viable CBMNC number was counted at certain intervals following expansion so as to screen the cytokines combination with the best expansion effect.Results On the day 14 of culture the best expansion effect was found in the group of TPO and SCF in which the number of CD61+ cells and viable CBMNC increased 48.40±5.71-fold and 3.14±0.62-fold respectively.The results were remarkably more than those of TPO along group(18.72±3.73-fold and 1.81±0.36-fold respectively.The number of expanded CD61+ cells in the combination of TPO and IL-3 increased 25.82±3.88-fold,but the percentage of CD61+ cells after expansion was only 9.34%±1.92%, which was less than that in TPO along group.No significant difference for expansion effect was observed between the groups of TPO along and the combination of TPO with EPO or SCGF.Conclusion SCF could synergize with TPO to expand megakaryocytes from CBMNC,and the combination of TPO and SCF significantly increases the number and purity of megakaryocytes in culture system.This combination of cytokines might provide experimental evidences for the study on megakaryocytopoiesis.