摘要
目的 探讨构建BCR-ABL逆转录病毒介导的小鼠慢性粒细胞白血病模型最佳实验条件,以简化实验过程、节约成本并提高建模成功率.方法 不同剂量5-氟脲嘧啶（5-FU）注射受体鼠后观察骨髓中原始细胞占有核细胞百分比;纤维连接蛋白（FN）铺板后观察逆转录病毒感染效率的变化;分别用不同数量的骨髓细胞（0.2～0.6）×106个进行移植后通过形态学观察和荧光原位杂交技术（fluorescence in situ hybridization,FISH）鉴定移植小鼠成模状况.结果 研究剂量范围内5-FU量不影响骨髓原始细胞相对数量;纤维连接蛋白铺板后可使活细胞数量增加30%～50%,感染效率提高10%～20%;移植0.5×106个成功感染了BCR-ABL逆转录病毒的供体鼠骨髓细胞入受体小鼠即可成功建模.结论 优化后的实验条件简化了程序,节约了成本,大大提高了建模成功率,为相关研究提供了良好的实验参考.
Objective To explore the optimal experimental conditions for the construction of the BCR-ABL retrovirus-mediated mouse chronic myeloid leukemia（CML）model,so that the experimental procedures can be simplified with less expenditure and elevated rate of success in leukemia model construction.Methods 5-FU was injected into the recipient mice at various dosages and the ratio of blast cells in all the nucleated cell in bone marrows were accounted;the retroviral infection efficiencies were evaluated in different conditions with or without fibronectin pre-plating;Morphological observation and in situ hybridization techniques were employed to identify the modeling status of the recipient mice after transplantation of donor bone marrow cells at various dosages （0.2×106～0.6×106 per mice）.Results 5-FU dosage within the reported range showed no significant effect on the relative ratio of bone marrow blast cell;Pre-plating with fibronectin led to a 30%～50% increase in the percentage of live cells and an enhancement of infection efficiency for 10%～20%;Transplantation of 0.5× 106 BCR-ABL retrovirus-infected donor bone marrow cells led to successful construction of the CML model in the recipient mice.Conclusion The optimized experimental conditions has simplified experimental procedures with lower expenditure and elevated rate of success in the construction of CML model mice,which has also provided an excellent reference for future relevant studies.
出处
《国际检验医学杂志》
CAS
2010年第6期526-527,530,共3页
International Journal of Laboratory Medicine
基金
深圳市科技计划项目资助项目(200903090)
关键词
白血病
髓样
慢性
纤连蛋白类
原位杂交
小鼠
模型
动物
leukemia,myelogenous,chronic
fibronectins
in situ hybridization
mice
models,animal
