%0 Journal Article
%T Production of u-PA with rCHO Cell Culture on Porous Microcarriers in Serum-free Growth Medium<br>多孔微载体无血清培养rCHO细胞生产u-PA
%A HU Xian Wen
%A XIAO Cheng Zu
%A LI Zuo Hu
%A GUO Zhi Xia GAO Li Hua
%A ZHANG Zheng Guang
%A XU Zhao Ping
%A WANG Fei
%A <br>胡显文
%A 肖成祖
%A 李佐虎
%A 郭志霞
%A 高丽华
%A 张正光
%A 胥照平
%A 王菲
%J 生物工程学报
%D 2000
%I 
%X A novel technique was developed to deal with apoptosis in large-scale animal cell culture. By means of replacing part of Cytopore porous microcarriers at regular intervals, a rCHO cell line, which produces urokinase-type plasminogen activitor(u-PA), was cultivated continuously with serum-free medium in a 30 L stirred tank for 91 days. The cell density was maintained at (1.3-2.6) x 10(7)/mL, and > 90% of cells was viable. In order to reduce the effect of cell density on cell growth and expression, a cyclic pressure oscillation was exerted on a 7.5 L reactor headspace to enhance cell expression at high cell density to a certain extent. During the 67 days of medium-replacement culture, the maximal cell density reached 2.64 x 10(7)/mL, and cell viability was always kept above 95% when combined with microcarrier-replacement. Compare to control culture, culture with cyclic pressure oscillation could enhance cell expression level and reduce the ratio of glucose metabolized anaerobically to produce lactate. With four-step purification process, about 80 g u-PA(approximately 90% scu-PA) was recovered from approximately 2100 liters supernatant which contained approximately 135 g u-PA.
%K Animal cell culture
%K porous microcarrier
%K prourokinase
%K serum
%K free medium
%K periodic artificial stimulus<br>动物细胞培养
%K 微载体更新
%K u－PA
%K rCHO细胞
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=425514EE358B5887&yid=9806D0D4EAA9BED3&vid=7801E6FC5AE9020C&iid=38B194292C032A66&sid=117BC32987199759&eid=AFB2C35009D5D191&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=13&reference_num=15