摘要
目的：以小鼠C2C12成肌细胞为对象开展血友病乙基因治疗研究。方法：首先将构建好的带有MCK增强手和hCMV启动子的反转录病毒载体G1NaMCⅨ转染PA317细胞，并用其上情感染C2C12细胞，所得克隆细胞在离体分别进行了Southern、Northern、Western鉴定。进一步用带有hFⅨcDNA的C2C12细胞进行C3H小鼠活体表达研究。结果：hFⅨ活体表达量与细胞注射数量、细胞离体表达量呈正相关，使用免疫抑制剂可提高hFⅨ活体表达量并延长表达时间；较长时间间隔后进行重复注射亦可延长hFⅨ表达时间。结论：同种异体成肌细胞存在免疫排斥现象，较长时间间隔重复注射可提高移植细胞存活率，从而有助于实现成肌细胞介导的血友病乙基因治疗。
Objective:To develop experimentally gene therapy for hemophilia B by using C2C12 cells and C3H mice as models. Methods :The retroviral vector containing MCK enhancer and hCMV promoter was constructed and was used to trans fect PA317 cells. Then the viral particles were used to trans fect C2C12 cells and select the G418r clones,which were further confirmed by Southern, Northern, Western blots. And then the C2C12 cells expressing hF Ⅸ protein were injected into C3H mice and hF a expression in vivo was studied. Results:The expression level of hF Ⅸ in vivo was positively correlated with the num ber of injected cells,and the in vitro expression level of these cell clones. The expression of hF Ⅸ could be enbanced and prolonged by immunosuppressive agents. Conclusion: Allogeneic myoblasts transplantation leads to immunorejection,but repeated injections at a longer intervals could improve the existing rate of transplanted cells. And this might be helpful to gain insight into the myoblast-mediated gene therapy for bemophilia B patients.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
1997年第9期471-473,共3页
Chinese Journal of Hematology
基金
国家"863"高技术重大项目基金!863-102-17-(2)
上海市科技发展基金!95JC14009
