IL-3影响脐血单个核细胞体外扩增、凋亡及再植入能力的研究

目的探讨细胞因子IL-3对脐血单个核细胞(MNC)体外扩增、凋亡和植入能力的影响,寻找扩增后脐血应用于临床移植的可行方案。方法采用早期作用的细胞因子组合对脐血MNC进行短期无血清悬浮培养,比较有或无IL-3支持的扩增效果和细胞凋亡变化;MNC扩增6d后移植给亚致死剂量照射的非肥胖糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠,观察6周后存活小鼠的人脐血细胞植入情况。结果脐血MNC在有IL-3支持的体系中培养6~10d获得最佳扩增效果,同时细胞表面膜联蛋白V(annexinⅤ)的表达明显减少;移植6周后存活小鼠的骨髓、脾和胸腺细胞中能检出人类CD45抗原,外周血提取的DNA也能检出人特异的Alu序列。结论短期培养下IL-3有助于脐血单个核细胞体外适度扩增,减少凋亡且并不损伤扩增后细胞的植入能力。

Effect of IL-3 on ex-vivo expansion, apoptosis and reimplantation of human cord blood mononuclear cells

Objective To study the effect of cytokine IL-3 on ex-vivo expansion, apoptosis and implantation capability of UCB MNCs, and to find a feasible way to apply the ex-vivo expanded UCB MNCs to clinical transplantation. Methods UCB MNCs were cultured for short-term in serum-free medium with early acting cytokine combinations, with or without IL-3. After 6-day culture, MNCs were transplanted into NOD/SCID mice, which were given sublethal dose irradiation. Implantation were observed after 6 weeks. Results UCB MNCs reached the best expansion between d6 and d10 with the addition of IL-3, and Annexin V expression on cell surface decreased obviously. Six weeks after transplantation, human CD45 antigen were detected by FCM on BM, spleen, and thymus cells, and the human specific Alu repetitive sequence was detected in the DNA obtained from peripheral blood of the survival mice. Conclusion In short-term culture, IL-3 increases the expansion of UCB MNCs, and decreases cell apoptosis but does not impair the implantation capability of expanded cells at the same time.