表达内皮抑素的rCHO细胞的微囊化培养  

Culture of Microencapsulated Endostatin-CHO Cells

作　　者：张英[1,2] 王为[1] 吕国军[1,2] 于炜婷[1] 郭昕[1] 雄鹰[1] 马小军[1] 

机构地区：[1]中国科学院大连化学物理研究所 [2]中国科学院研究生院北京100039

出　　处：《中国生物工程杂志》2007年第9期1-7,共7页China Biotechnology

基　　金：国家自然科学基金重点项目(20236040);国家"973"计划资助项目(2005CB522702);辽宁省自然科学基金资助项目(20062149)

摘　　要：微囊化重组细胞移植治疗肿瘤是一种新兴的肿瘤基因治疗方法,如果将此技术应用到临床研究,就需要制备大量的细胞活性良好、重组蛋白表达量高的生物微胶囊。种子细胞是生物微胶囊治疗作用的执行者,是构建微囊微反应器的基本元素。如何获得大量高活性的种子细胞已经成为规模化制备生物微胶囊所面临的关键的限制因素。考察了搅拌式生物反应器内扩增的rCHO细胞进行包囊及微囊化细胞在生物反应器内规模化培养的可行性。实验结果显示:rCHO细胞在生物反应器内可以快速生长,并且对数期细胞包囊,微囊化细胞活性良好。制备的微囊化细胞可以在生物反应器内培养,与培养板培养比较细胞生长较快、内皮抑素表达量较高。应用生物反应器培养技术能够在体外快速、大量扩增rCHO细胞,满足微囊化细胞制备对种子细胞量与质的要求,微囊化细胞可以在生物反应器内培养。Microencapsulated recombinant cells technology is a novel approach to tumors therapy. It is great importance in encapsulated cells therapy to find a proper way for acquiring seed cells in large-scale in a short time for large scale preparation of biological microcapsule with high cell viability and recombinant protein production, The way and technique to gain adequate seed cells for cell encapsulation with bioreactor culture system were explored. The results showed that the recombinant CHO cells could expand rapidly in the bioreactor in three dimensions and the cell amount reached about 20 times of the initial amount after 10 days culture. The expanded cells could be encapsulated into the microcapsule and the microencapsulated cells could keep high viability and expand rapidly in the bioreactor. In comparison with the cells statically cultured in 24-well plates, a quicker growth and a higher endostatin production of microencapsulated cells were observed. B. Braun bioreactor provided the culture environment with relatively low shear force and good communication among cells in threedimension, therefore recombinant CHO cells showed a strong ability to expanded in vitro. Large number of seed cells can be obtained rapidly with this culture system, which can meet the requirement of encapsulated cells therapy in both quantity and quality.

关 键 词：微胶囊 种子细胞培养 生物反应器 规模化培养rCHO细胞 

分 类 号：Q813.1[生物学—生物工程]