体外扩增的脐血细胞移植BABL/c小鼠的实验研究

Study of Transplanting Human Fresh and Expanded Cord Blood Cells into BABL/c Mice

目的探讨体外扩增后脐血单个核细胞(CBMC)的应用价值。方法随机分组的BABL/c小鼠经亚致死量照射后,每组分别由尾静脉注入新鲜CBMC悬液、不同细胞因子组合(TPO+FL+IL-6、TPO+FL+IL-6+IL-3)扩增后的CBMC悬液、脐血全血和生理盐水。比较各组小鼠的存活率及造血恢复情况,并应用PCR和免疫组化方法观察小鼠体内人源细胞植入情况。结果扩增前后CBMC均可在BABL/c小鼠体内植入并重建造血功能,且存活率与植入率无显著性差异(P>0.05)。实验组造血恢复情况优于对照组。含IL-3的细胞因子组合对CBMC的扩增效果(8.96±0.52倍)优于不含IL-3者(6.28±0.16倍)(P<0.01)。结论体外扩增后CBMC数量显著增加,且保持了其重建造血功能及植入潜能,比新鲜的CBMC有更大的应用价值。

Objective To evaluate the applicability of cord blood mononuclear cells（CBMC） expanded in vitro by analyzing the engraftment sign and the hematopoiesis reconstitution of BABL/c mice after they were infused using fresh or explanded CBMC. Methods Sublethally irradiated BABL/c mice were randomly divided to receive either fresh CBMC,or CBMC cultured with two different cytokine combinations（TPO ＋ FL＋IL-6 and TPO ＋ FL＋IL-6＋IL-3）,or cord blood,or normal saline respectively. The survival rates and hematopoiesis reconstitutions were observed and compared among them. The evidence of human hematopoiesls was analyzed by polymerase chain reaction（PCR）and immunohistochemistry. Results Both fresh and expanded CBMC could be implanted into BABL/c mice to produce donor-recipient chimera and to reconstitute hematopoiesis,showing no significant differences in survival rate and engrafting rate between the two groups（P〉0.05）. As for hematopolesis reconstltution,the data collected from trans- fused mice were significantly higher than those from control group. The proliferative effect of cytoklne combination including IL-3 （8.96 ±0.52 floded） was higher than the other one（6.28 ± 0.16 floded, P 〈 0.01）. Conclusion The cytokine combinations could significantly increase CBMC in vitro. Expanded CBMC could keep their engraftment potential and ability to reconstitute hematopoietic system.