人间充质干细胞转染条件的优化  

Condition Optimization of Human Mesenchymal Stem Cells Transfection

作　　者：李云霞 边延林 朱建伟 张宝红 LI Yunxia;BIAN Yanlin;ZHU Jianwei;ZHANG Baohong(Engineering Research Center of Cell&Therapeutic Antibody(MOE),School of Pharmacy,Shanghai Jiao Tong University,Shanghai 200240)

机构地区：[1]上海交通大学药学院,细胞工程及抗体药物教育部工程研究中心,上海200240

出　　处：《中国医药工业杂志》2023年第5期742-752,共11页Chinese Journal of Pharmaceuticals

摘　　要：间充质干细胞(MSCs)在人体内分布广泛,具有强大的自我更新能力、多能性以及免疫调节能力,并且具有迁移到受损组织和肿瘤部位的特性,表达抗体的MSCs在肿瘤治疗方面显示出良好的应用前景。然而,MSCs难以转染,限制了实际应用。该文采用4种方法转染脐带血来源的人间充质干细胞,并优化了转染条件。结果表明,三质粒系统制备慢病毒转染MSCs的效率较高,当包装条件为m(DNA)∶m(Lipo)=1∶2、m(vector)∶m(psPAX2)∶m(VSVG)=3∶2∶1时,50%的MSCs可成功表达目的蛋白。利用优化后的条件,本研究成功构建了分别表达Anti-CD24和BiTE-Fc 2种不同结构抗体的MSCs,为MSCs在肿瘤治疗领域的应用研究提供了试验基础。Mesenchymal stem cells(MSCs)are distributed widely in the human body.MSCs have powerful selfrenewal ability,pluripotency,and immunomodulation property.Due to the damaged tissue and tumor site tropism,MSCs that express antibodies have potential applications in tumor therapy.However,the low efficiency of MSC transfection restricts their application.This study optimized four transfection methods for human umbilical cord blood-derived MSCs.The results showed that with the optimal packaging conditions[m(DNA)∶m(Lipo)=1∶2,m(vector)∶m(psPAX2)∶m(VSVG)=3∶2∶1],lentivirus produced by three-plasmid system could infect MSCs more efficiently,50%of MSCs could express target protein successfully.In addition,MSCs expressing two antibodies with different structures,Anti-CD24 and BiTE-Fc,were successfully constructed with the above methods,respectively,providing the research basis for applying MSCs in tumor therapy.

关 键 词：间充质干细胞 抗体表达 慢病毒 转染 

分 类 号：Q816[生物学—生物工程]