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机构地区:[1]成都军区昆明总医院血液科 [2]南方医科大学南方医院血液科,广州,510515
出 处:《白血病.淋巴瘤》2008年第2期-,共3页Journal of Leukemia & Lymphoma
基 金:国家自然科学基金
摘 要:目的 观察经rAAV-2-hGM-CSF、rAAV-2-mGM-CSF基因转导修饰的骨髓间充质干细胞在体基因表达情况.方法 按预先摸索好的转染条件在体外用rAAV-2-hGM-CSF、rAAV-2-mGM-CSF感染骨髓间充质干细胞后,继续在体外扩增培养12 d,再将被基因修饰的骨髓间充质干细胞通过尾静脉回输到6周龄的裸鼠体内,对照组回输未经基因转导处理的骨髓间充质干细胞,分别于此后的2、4、6、8周处死裸鼠,取其外周血,计数白细胞总数,检测血清中hGM-CSF、mGM-CSF的含量.结果 回输前hGM-CSF、mGM-CSF的分泌水平为36.25、25.14 ng/L,回输后2、4、6、8周裸鼠血清中hGM-CSF的含量分别为23.77、26.32、19.77、15.25 ng/L,mGM-CSF的含量分别为34.96、34.84、35.50、32.93 ng/L,相应时间点对照组裸鼠血清中mGM-CSF的水平分别为17.34、17.44、14.68、16.85 ng/L.rAAV-2-mGM-CSF转导组可使裸鼠体内白细胞计数增加,而rAAV-2-hGM-CSF转导组和对照组裸鼠体内的白细胞计数却无变化.结论 骨髓间充质干细胞可作为一种基因治疗的载体细胞,携带在体外经过基因修饰了的治疗基因在体内发挥其治疗作用.Objective To observe the in vivo gene expression profile of the recombinant adenoassociated-2 virus mediated human GM-CSF, mouse GM-CSF (rAAV-2-hGM-CSF, rAAV-2-mGM-CSE )vector modified bone marrow mesenchymal stem cell (BMSC). Methods We transduced the BMSC by rAAV-2-hGM-CSF, rAAV-2-mGM-CSF at the condition which have acquired before respectively, then transfused the in vitro gene modified BMSC after 12 days proliferation in vitro to 6 weeks old nude mice through tail vein,while the BMSC transfused in control group hadn' t been gene modified. 2, 4, 6, 8 weeks after transfusion, count the total white blood cells and detect the hGM-CSF, mGM-CSF concentration in nude mice serum at that time point. Results Nude mice serum hGM-CSF levels were 23.77, 25.32, 19.77, 15.25 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 36.25 ng/L, the in vitro level before transfusion; mGM-CSF levels were 34.96, 34.84, 35.50, 32.93 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 25.14 ng/L, the in vitro level before transfusion; at the same time point the nude mice serum mGM-CSF levels were 17.34,17.44, 14.68, 16.85 ng/L in control group, rAAV-2-mGM-CSF transduced BMSC made the nude mice white blood cell count increased, but no changes in nude mice white blood cell count at rAAV-2-hGM-CSFtransduced BMSC and control group. Conclusion BMSC as a gene therapy vehicle, it can be gene modified in vitro, then the gene modified BMSC could let the therapeutic gene to have therapeutic effects in vivo.
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