磁性搅拌珠制备及在NK-92细胞体外扩增中的应用  

Preparation of magnetic stirring beads and its application in NK-92 in vitro expansion

作　　者：刘洋洋 周乐豪 郝梦洋 蔡海波[1] 谭文松[1] LIU Yang-yang;ZHOU Le-hao;HAO Meng-yang;CAI Hai-bo;TAN Wen-song(State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《高校化学工程学报》2021年第4期683-689,共7页Journal of Chemical Engineering of Chinese Universities

摘　　要：悬浮细胞的生长对剪切力较敏感,在搅拌式生物反应器中采用球体搅拌可减少流体剪切力对细胞扩增的影响。为此,研究采用两步法制备海藻酸钠/壳聚糖双组分磁性搅拌珠并将其运用于磁力搅拌瓶,用于NK-92细胞的动态培养。结果显示,当海藻酸钠质量浓度达到25 mg×mL^(-1)时,磁性搅拌珠的力学性能最佳;用于NK-92细胞扩增时,在保证细胞活性的同时,总细胞扩增倍数以及扩增后的细胞杀伤活性分别可达到(72.63±7.80)倍和(85.57±3.69)%,显著高于T 25培养瓶静态培养时的(22.41±1.46)倍(p<0.05)和(70.53±1.72)%(p<0.05)。研究结果为悬浮细胞动态培养设备的设计提供了新的思路,为免疫细胞的体外扩增提供了技术支持。Spherical type agitated bioreactors with low shearing force are generally applied in expanding suspension cells.In this study,sodium alginate/chitosan two-component magnetic stirring beads were prepared by a two-step method for dynamic culture of NK-92 cells.The results show that the best mechanical properties of the magnetic stirring beads are those prepared with 25 mg×mL^(-1) sodium alginate.The total cell expansion of NK-92 cells under dynamic culture conditions is(72.63±7.80)folds,which is significantly higher than that prepared with T 25 flask((22.41±1.46)folds,p<0.05).The killing activity of the expanded cells under dynamic culture conditions also increased significantly(85.57±3.69)%when compared with T 25 flask(70.53±1.72)%,(p<0.05).This study provides promising results for in vitro expansion of immune cells and dynamic culture development.

关 键 词：NK-92细胞 磁性搅拌珠 海藻酸钠 体外扩增 生物反应器 

分 类 号：Q2-33[生物学—细胞生物学]