Summary
Three monkey kidney cell lines and primary chicken embryo cells were grown in microcarrier culture. The carrier support was DEAE-Sephadex gel beads at low anion exchange capacity prepared according to a protocol developed at the Massachusetts Institute of Technology.
The growth rate of the cells and the final cell density in microcarrier culture was dependent on the concentration of the beads in culture and on the size of the initial cell inoculum. A bead concentration of 1.0 to 2.0 mg of beads/ml of tissue culture medium and a cell inoculum of 20,000 cells/cm2 of bead surface appeared to be optimal. The efficiency of the microcarrier culture system was compared to that of stationary and roller bottle cultures. Stationary flasks gave cell densities about twofold higher than maximal densities in roller bottles and about threefold and twofold higher than cell densities in microcarrier culture at a bead concentration of 2.5 and 1.0 mg/ml, respectively.
In terms of cell yield per millitier of tissue culture medium, the microcarrier culture was superior to roller bottle and stationary cultures. An advantage of the microcarrier culture system is its suitability for a scale up into large volume production units.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Van Wezel, A. L.. Growth of cell strains and primary cells on microcarriers on homogeneous culture. Nature 216: 64–65; 1967.
Van Hemert, P.; Kilburn, D. G.: Van Wezel, A. L. Homogeneous cultivation of animal cells for the production of virus and virus products. Biotechnol. Bioeng. 11: 875–885; 1969.
Van Wezel, A. L. Microcarrier cultures of animal cells. Kruse, P. F.; Patterson, M. K., Jr. eds. Methods and applications of tissue culture. New York: Academic Press; 1973: 372–377.
Van Wezel, A. L.; van der Velden-de-Groot, C. A. M. large scale cultivation of animal cells in microcarrier culture. Process Biochemistry 13: 6–8; 1978.
Van Wezel, A. L. New trends in the preparation of cell substrates for the production of virus vaccines. Prog. Immunobiol. Stand. 5: 187–192; 1972.
Van Wezel, A. L.; Van Steenis, G.; Hannik, C. A.; Cohen, H. New approach to the production of concentrated and purified inactivated polio and rabies tissue culture vaccines. Dev. Biol. Stand. 41: 159–168; 1978.
Van Wezel, A. L.; Van Steenis, G.. Production of an inactivated rabies vaccine in primary dog kidney cells. Dev. Biol. Stand. 40: 69–75; 1977.
Meignier, B.. Cell culture on beads used for the industrial production of foot and mouth disease virus. Dev. Biol. Stand. 42: 141–145; 1978.
Spier, R. E.; Whiteside, J. P. The production of foot and mouth disease virus from BHK21C13 cells grown on the surface of DEAE-Sephadex A50 beads. Biotechnol. Bioeng. 18: 659–667; 1976.
Giard, D. J.; Thilly, W. G.; Wang, D. I. C.; Levine, D. W. Virus production with newly developed microcarrier system. Appl. Environ. Microbiol. 34: 668–672; 1977.
Girard, D. J.; Loeb, D. H.; Thilly, W. G.; Wang, D. I. C.; Levine, D. W. Human interferon production with diploid fibroblast cells grown on microcarriers. Biotechnol. Bioeng. 21: 443–442; 1979.
Levine, D. W.; Wong, J. S.; Wang, D. I. C.; Thilly, W. G. Microcarrier cell culture: new methods for research-scale applications. Somatic Cell Res. 3: 149–155; 1977.
Levine, D. W.; Wang, D. I. G.; Thilly, W. G. Optimizing parameters for growth of anchorage dependent mammalian cells in microcarrier culture Acton, R. T.; Lynn, J. D. eds. Cell culture and its application. New York: Academic Press; 1977: 191–216.
Levine, D. W.; Thilly, W. G.; Wang, D. I. C. Treatment of cell culture microcarriers. US Patent No. 4.036.693; 1974.
Thilly, W. G.; Levine, D. W. Microcarrier culture: a homogeneous environment for studies of cellular biochemistry. Jacoby, W. B. ed. Methods in enzymology. New York: Academic Press; 1979: 184–194.
Levine, D. W. Production of anchorage dependent mammalian cells on microcarriers. Cambridge, Massachusetts: Department of Nutrition and Food Science, Massachusetts Institute of Technology; 1976. Thesis.
Yasumura, Y.; Kawakita, Y. Research into SV40 by tissue culture. Nippon Rinsho, 21: 1201–1205; 1963.
Hull, R. N.; Cherry, W. R.; Johnson, I. S. The adaptation and maintenane of mammalian cells to continuous growth in tissue culture. Anat. Rec 124: 490–492; 1956.
Jensen, F. C.; Girardi, A. J.; Gilden, R. V.; Koprowski, H. Infection of human and simian tissue cultures with rous sarcoma virus. Proc. Natl. Acad. Sci. USA 52: 53–59; 1964.
Rubin, H. 1973. Preparation of primary chick embryo cells. Kruse, P. F.; Patterson, M. K., Jr. eds. Tissue culture methods and applications. New York: Academic Press; 1973: 119–122.
Van Wezel, A. L. The large scale cultivation of diploid cell strains in microcarrier culture: improvement of microcarriers. Dev. Biol. Stands. 37: 143–147; 1973.
Horng, C. B.; McLimans, W. F. Primary suspension culture of calf anterior pituitary cells on a microcarrier surface. Biotechnol. Bioeng. 17: 713–732; 1975.
McLimans, W. F. Mass culture of mammalian cells. Jacoby, W. B.; ed. Methods in enzymology. Vol. LVIII. New York: Academic Press; 1979: 194–211.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mered, B., Albrecht, P. & Hopps, H.E. Cell growth optimization in microcarrier culture. In Vitro 16, 859–865 (1980). https://doi.org/10.1007/BF02619423
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02619423