人脐血单个核细胞体外扩增后植入NOD/SCID小鼠重建多系造血  

Implantation of ex vivo expanded human cord blood mononuclear cells into NOD/SCID mice

作　　者：毛平[1] 彭盘俐[1] 许力[1] 

机构地区：[1]广州医学院附属广州市第一人民医院血液内科,510180

出　　处：《中华器官移植杂志》2006年第4期231-234,共4页Chinese Journal of Organ Transplantation

基　　金：广东省科技计划基金资助项目(B30202);广州市科学技术局科技攻关重点项目基金资助项目(2002z2-E0241)

摘　　要：目的探讨人脐血单个核细胞（MNC）体外扩增后植入能力的改变，以及对NOD／SCID小鼠造血重建的影响。方法采用不同细胞因子组合对人脐血MNC进行短期无血清培养扩增，比较扩增后的效果及细胞凋亡的变化；将扩增6d后的人脐血MNC植入经半致死剂量照射的NOD／SCID小鼠体内，观察小鼠6周后的存活率和造血重建情况。结果人脐血MNC在干细胞因子（SCF）、酪氨酸激酶受体3配基（FL）、白细胞介素6（IL-6）和白细胞介素3（IL-3）细胞因子共同作用下，培养6～11）d获得最佳扩增效果，同时细胞表面膜联蛋白V（Annxin V）的表达明显减少；移植6周后有55．6％的小鼠存活，存活小鼠的骨髓、脾和胸腺细胞中均能检测出人类CD45、CD34、CD33、CD3和CDl9抗原，外周血提取的DNA可检测出人特异的Cart—Ⅰ基因和Alu基因。结论SCF＋FL＋IL-6＋IL-3细胞因子协同作用能有效扩增人脐血MNC，MNC扩增6d后可成功植入N（）D／SCID小鼠体内，并帮助小鼠重建多系造血。Objective To elucidate the effect of ex vivo expansion of umbilical cord blood （UCB） mononuclear cells （MNCs） on their implantation capability and the hematopoietic reconstitu tion, and to find a feasible way of applying ex vivo expanded UCB MNCs to clinical transplantation. Methods UCB MNCs were cultured in short-term in serum-free medium with different early acting cytokines combinations in order to observe the amplification result and cells apoptotic difference. The 6-day expanded cells were transplanted into sublethally irradiated NOD/SCID mice to assess the implantation and the hematopoietic reconstitution of the survival mice. Results UCB MNCs reached the best amplification result between day 6 and day 10 with the contribution of SCF, FL, IL-6 and IL-3 in common and the presence of Annxin V on the surface of cells obviously decreased. Six weeks after transplantation, CD45, CD34, CD33, CD3 and CD19 antigen could be detected by FCM on BM, spleen and thymus cells of the alive mice and the human specific Alu and Cart-Ⅰ repetitive sequence could be detected in the DNA obtained from peripheral blood by PCR. Conclusion After 6 day effective expanding with SCF＋ FL ＋ IL-6 ＋ IL-3 UCB MNCs can be implanted into NOD/SCID mice suc cessfully and contribute to reconstitute the multiple hematopoiesis.

关 键 词：脐血干细胞移植 人类 小鼠 

分 类 号：R457.7[医药卫生—治疗学]