CAJ | 학술논문

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.
목적 관찰경rAAV-2-hGM-CSF、rAAV-2-mGM-CSF기인전도수식적골수간충질간세포재체기인표체정황.방법 안예선모색호적전염조건재체외용rAAV-2-hGM-CSF、rAAV-2-mGM-CSF감염골수간충질간세포후,계속재체외확증배양12 d,재장피기인수식적골수간충질간세포통과미정맥회수도6주령적라서체내,대조조회수미경기인전도처리적골수간충질간세포,분별우차후적2、4、6、8주처사라서,취기외주혈,계수백세포총수,검측혈청중hGM-CSF、mGM-CSF적함량.결과 회수전hGM-CSF、mGM-CSF적분비수평위36.25、25.14 ng/L,회수후2、4、6、8주라서혈청중hGM-CSF적함량분별위23.77、26.32、19.77、15.25 ng/L,mGM-CSF적함량분별위34.96、34.84、35.50、32.93 ng/L,상응시간점대조조라서혈청중mGM-CSF적수평분별위17.34、17.44、14.68、16.85 ng/L.rAAV-2-mGM-CSF전도조가사라서체내백세포계수증가,이rAAV-2-hGM-CSF전도조화대조조라서체내적백세포계수각무변화.결론 골수간충질간세포가작위일충기인치료적재체세포,휴대재체외경과기인수식료적치료기인재체내발휘기치료작용.
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.