Abstract
As part of an effort to broaden the applicability and efficiency of microcarrier cell culture various alternative new microcarriers were synthesized. The microcarriers were compared as substrates for the growth of several types of cells and with respect to binding of proteins from the culture medium. Crosslinked dextran has been found to be the most suitable material for a microcarrier matrix and was used as the matrix for the new microcarriers. One type of microcarrier was synthesized so that the charges necessary for cell attachment were present only in the surface layer of the microcarrier in the form of N,N,N-trimethyl-2-hydroxyaminopropyl groups. The resulting microcarriers had a very low capacity to bind proteins from the culture medium (e.g. albumin and IgG) and such proteins could be removed from the cultures more efficiently than when using previous microcarriers. A new principle was used for the development of the other type of microcarrier. A surface layer of cross-linked denatured collagen provided the surface for growth of cells. These microcarriers provided a “natural” substrate for cell growth and allowed improved attachment and spreading of cells with epithelial morphology. Harvesting cells from these microcarriers with proteolytic enzymes was more efficient than with previous microcarriers. The two alternative microcarriers should result in improved process efficiency and have a potential value in the preparation of live vaccines.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
P. Bornstein and H. Sage, 1980, Structurally distinct collagen types, Ann. Rev. Biochem., 49:957–1003.
E. Engvall and E. Rouslahti, 1977, Binding of soluble form of fibroblast surface protein, fibronectin to collagen, Int. J. Cancer, 20:1–5.
E. Engvall, E. Rouslahti and E.J. Miller, 1978, Affinity of fibronectin to collagens of different genetic types and to fibrinogen, J. Exp. Med., 147:1584–1595.
L.I. Gold and E. Pearlstein, 1980, Fibronectin-collagen binding and requirements during cellular adhesion, Biochem. J., 186:551–559.
D.J. Giard and R.J. Fleischaker, 1980, Examination of parameters affecting human interferon production with microcarrier-grown fibroblast cells, Antimicrobial Agents Chemotherap., 18:130–136.
F. Grinnell, 1978, Cellular adhesiveness and extra-cellular substata, Int. Rev. Cytol., 53:65–144.
F. Grinnell, M. Feld and D. Minter, 1980, Fibroblast adhesion to fibronogen and fibrin substrata: Requirement for cold insoluble globulin (plasma fibronectin), Cell, 19:517–525.
F. Gubenŝek and S. Lapanje, 1968, Potentiometric titration studies of diethylaminoethyl dextran base, J. Macromol. Sci. Chem., A2:1045–1054.
M. Hirtenstein and J. Clark, 1980, Attachment and proliferation of animal cells on microcarriers (Cytodex ™1). In “Tissue Culture in Medical Research” eds. R.J. Richards, K.T. Rajan, pp.97–104, Pergamon: Oxford.
M. Hirtenstein, J. Clark, G. Lindren and P. Vretblad, 1980, Microcarriers for animal cell culture: a brief review of theory and practice, Develop. Biol. Standard, 46:109–116.
M. Höök, K. Rubin, Å. Oldenberg, B. Öbrink and A. Vaheri, 1977, Cold-insoluble globulin mediates the adhesion of rat liver cells to plastic petri dishes, Biochem. Biophys. Res. Coram., 79: 726–733.
D.W. Levine, D.I.C. Wang and W.G. Thilly, 1979, Optimization of growth surface parameters in microcarrier culture, Biotechnol. Bioeng., 21:821–845.
M. Manousos, M. Ahmed, C. Torchio, J. Wolff, G. Shibley, R. Stephens and S. Mayyasi, 1980, Feasibility studies of oncornavirus production in microcarrier cultures, In Vitro, 16:507–515.
B. Meignier, H. Mougeot and H. Favreè, 1980, Foot and mouth disease virus production on microcarrier-grown cells, Develop. Biol. Standard, 46:249–256.
B. Mered, P. Albrecht and H.E. Hopps, 1980, Cell growth optimization in microcarrier culture, In Vitro, 16:859–865.
B.J. Montagnon, B. Fanget and A.J. Nicolas, 1981, The largescale cultivation of Vero cells in microcarrier culture for virus vaccine production. Preliminary results for killed poliovirus vaccine, Develop. Biol. Standard., 47: 55–64.
Pharmacia Fine Chemicals, 1980, Microcarrier cell culture, Technical notes, 1, Uppsala, Sweden.
Pharmacia Fine Chemicals, 1974, Dextran Fractions, Dextran Sulphate, DEAE-Dextran. Defined polymers for biological research, Technical Book Series, Uppsala, Sweden.
L.M. Reid and M. Rojkind, 1979, New techniques for culturing differentiated cells: reconstituted basement membrane rafts, Methods. Enzymol., 58: 263–278.
K.K. Sanford, W.R. Earle, V.J. Evans, H.K. Waltz and J.E. Shannon, 1951, The measurement of proliferation in tissue cultures by enumeration of cell nuclei, J. Natl. Cancer Res. Inst., 11:773–795.
A, von Seefried and J.H. Chun, 1981, Serially subcultivated cells as substrates for poliovirus production for vaccine, Develop. Biol. Standard., 47:25–33.
A.L. van Wezel, C.A.M. van der Velden-de Groot and J.A.M. van Herwaarden, 1980, The production of inactivated polio vaccine on serially cultivated kidney cells from captive-bred monkeys, Develop. Biol. Standard., 46:151–158.
D. Yaffe, 1973, Rat skeletal muscle cells. In “Tissue Culture: Methods and Applications”, eds. P.F. Kruse, M.K. Patterson, pp. 106–114, Academic Press, New York.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1984 Plenum Press, New York
About this chapter
Cite this chapter
Gebb, C. et al. (1984). Alternative Surfaces for Microcarrier Culture of Animal Cells. In: Acton, R.T., Daniel Lynn, J. (eds) Eukaryotic Cell Cultures. Advances in Experimental Medicine and Biology, vol 172. Springer, New York, NY. https://doi.org/10.1007/978-1-4615-9376-8_9
Download citation
DOI: https://doi.org/10.1007/978-1-4615-9376-8_9
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4615-9378-2
Online ISBN: 978-1-4615-9376-8
eBook Packages: Springer Book Archive