Abstract
Using porous microcarrier Cytopore and a low-serum medium supplement BIGBEF-3, we have successfully cultivated recombinant CHO cell line CL-11G producing prourokinase and hybridomas producing anti-prourokinase monoclonal antibody in Celligen 1.5 or 5 L bioreactor. The cell density obtained ranged from 1 to 2 × 107 cells mL-1. The yields of prourokinase and monoclonal antibody increased with increasing cell density. As the cells could spontaneously release from and reattach to porous microcarriers, it was very easy to scale-up the cultivation. Thus the bead to bead cell transfer method has been used to scale up the cultivation of CL-11G cells to a 20 L reactor-scale for the pilot production of prourokinase, and also to scale-up the culture of hybridomas for the production of monoclonal antibody for the purification of prourokinase.
Similar content being viewed by others
References
Ai X, Ye J, Xiao C and Zhang Z (1995) Preparation and characterization of monoclonal antibodies against uPA and their application for purification of uPA. Bull Acad Mil Med Sci 19: 248-253.
Gotoh T, Honda H, Shiragami N and Unno H (1993) A new type porous carrier and its application to culture of suspension cells. Cytotechnol 11: 35-40.
Jia X and Xiao C (1993) MTT method for the estimation of the number of cells cultivated in microcapsules. Bull Acad Mil Med Sci 17: 207-211.
Kamiya K, Yanagido K and Shirokaze J (1995) Subculture for large scale cell culture using macroporous microcarrier. In: Beuvery EC, Griffiths JB and Zeijlemaker WP (eds) Animal Cell Technology: Development towards the 21st Century (pp. 759-763). Kluwer Acad Publishers, The Netherlands.
Li F, Li J, Li X, Lin J and Yu W (1993) High-level expression of pro-urokinase cDNA in Chinese hamster ovary cell line (1). Bull Acad Mil Med Sci 17: 89-92.
Looby D and Griffiths B (1990) Immobilization of animal cells in porous carrier culture. TIBTECH 8: 204-209.
Nilsson K, Buzsaky F and Masboch K (1986) Growth of anchorage-dependent cells on macroporous microcarriers. Bio/Technol 4: 989-990.
Reuveny S (1985) Microcarriers in cell culture: Structure and applications. Adv Cell Culture 4: 213-247.
Shirokaze J, Nogawa M and Ogura R (1994) High density culture using macroporous microcarrier. In: Spier RE and Griffith B (eds) Products for today, prospects for tomorrow (pp. 261-263). Oxford, Butterwerth-Heinemann.
Van Wezel AL (1967) Growth of cell-strains and primary cells on microcarriers in homogeneous culture. Nature 216: 64-65.
Xiao C, Zhou H and Huang Z (1994a) An autocontroller system for cell perfusion cultivation on microcarrier. Chinese Patent, ZL 91208893.1, 1994-09-14.
Xiao C, Huang Z, Liu F, Guo Z and Gao L (1994b) High density cultivation of genetically-engineered CHO cell lines with microcarrier culture systems. Chin Med Sci J 9: 71-74.
Xiao C, Huang Z, Zhang Z, Ye J, Gao L, Guo Z, Cheng D, Zhou H and Kong W (1994c) High density cultivation of a recombinant CD-1 cell line producing prourokinase using a Biosilon microcarrier culture system. Chin Med Sci J 9: 203-208.
Xiao C, Huang Z, Li W, Qu W, Gao L and Liu G (1996) High density cultivation of recombinant CHO cell line and two strains of hybridomas with porous microcarrier Cytopore. Bull Acad Mil Med Sci 20: 191-194.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Xiao, C., Huang, Z., Li, W. et al. High density and scale-up cultivation of recombinant CHO cell line and hybridomas with porous microcarrier Cytopore. Cytotechnology 30, 143–147 (1999). https://doi.org/10.1023/A:1008038609967
Issue Date:
DOI: https://doi.org/10.1023/A:1008038609967