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目的 :为了保护骨髓免受或少受化疗药物的损伤 ,用转染人类突变二氢叶酸还原酶 (DHFR)基因反转录病毒载体 ,研究其耐药特性及在小鼠造血细胞的表达和对造血细胞的保护作用。方法 :将含有DHFR基因的病毒上清转染小鼠骨髓造血细胞 ,将转染后的骨髓细胞从尾静脉回输给经致死量 (9Gy)照射的小鼠 ,用MTX和G4 18筛选。结果 :经G4 18和MTX筛选均可得到阳性克隆 ,实验组小鼠白细胞计数和红细胞压积逐渐恢复正常 ,而对照组小鼠 30d内全部死亡。结论 :经初步研究表明 ,该反转录病毒载体能成功转染小鼠造血细胞 ,并表达目的基因 ,使在MTX筛选下重建小鼠造血功能。
OBJECTIVE: To protect bone marrow from or to be damaged by chemotherapeutic agents, we used the retroviral vector transfected with the human mutant dihydrofolate reductase (DHFR) gene to study its drug resistance characteristics and its expression in mouse hematopoietic cells. Hematopoietic cell protection. METHODS: The viral supernatant containing the DHFR gene was transfected into mouse bone marrow hematopoietic cells. The transfected bone marrow cells were returned from the tail vein to lethally irradiated (9 Gy) irradiated mice and screened with MTX and G418. Results : Positive clones were obtained by G418 and MTX screening. The white blood cell count and hematocrit of mice in the experimental group gradually returned to normal, while mice in the control group died within 30 days. Conclusion: Preliminary studies have shown that the retroviral vector can successfully transfect mouse hematopoietic cells and express the target gene, so that the hematopoietic function of mice can be reconstructed under the MTX screen.