用改装的Ｓuper—Spinner搅拌瓶连续灌流培养产凝血酶原ＣＨＯ工程细胞

在动物细胞培养过程中对培养体系实施培基连续灌流能及时地补充细胞生长所需的营养物质、控制细胞代谢产物对细胞的影响 ,实现细胞的高密度长期培养 ,提高目的产品的生产效率1,2 ] 。细胞连续灌流培养的前提是在实施培基连续灌流的同时培养体系能有效地截留细胞3] 。这一前提增加了细胞培养装置的复杂程度 ,使之特化为价格昂贵的生物反应器 ,限制了细胞连续灌流培养的应用。如能通过对普通的细胞搅拌培养瓶进行改进 ,使之能用于细胞的连续灌流培养 ,则有利于细胞连续灌流培养的推广应用。1　材料和方法1 1　细胞产人重组凝血酶原ＣＨＯ工…

Continuously Perfused Cultivation of Genetically-engineered CHO Cells Producing Prothrombin in a Modified Super-Spinner

A Super-Spinner was Modified by mounting a stainless steel filter(pore size 75 microns) to the impeller shaft to retain cells while fresh nutrient is perfused. Using Macroporous microcarrier Cytopore 1, continuously perfused cultivation of a recombinant CHO cell line, CHO2DS producing prothrombin was performed with the perfusion of a protein-free medium DF6S. The cell retention rate was more than 90% during the 24 days continuously perfused cultivation. The viable cell density of CHO2DS and prothrombin concentration reached 4.62 x 10(6)(cells.m/L) and 11.3(mg/L) respectively after 9 days culture.