Production of u-PA with rCHO Cell Culture on Porous Microcarriers in Serum-free Growth Medium
10.3321/j.issn:1000-3061.2000.03.021
- VernacularTitle:多孔微载体无血清培养rCHO细胞生产u-PA
- Author:
Xian-Wen HU
1
;
Cheng-Zu XIAO
;
Zuo-Hu LI
;
Zhi-Xia GUO
;
Li-Hua GAO
;
Zheng-Guang ZHANG
;
Zhao-Ping XU
;
Fei WANG
Author Information
1. Institute of Biotechnology, Academy of Military Medical Sciences
- Keywords:
Animal cell culture;
porous microcarrier;
prourokinase;
serum-free medium;
periodic artificial stimulus
- From:
Chinese Journal of Biotechnology
2000;16(3):387-391
- CountryChina
- Language:Chinese
-
Abstract:
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 30L stirred tank for 91 days. The cell density was maintained at (1.3～2.6)×107/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 7.5L 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 2.64×107/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(～90 % scu-PA)was recovered from～2100 liters supernatant which contained～135 g u-PA.
