Summary
Plastic dishes were coated with an agarose layer. The layer was modified by covalently binding proteins to it, using the CNBr-method. Cells were seeded on the dishes and the number of attached cells was evaluated. The specificity of the attachment, was demonstrated by showing that cells, carrying specific membrane-bound immunoglobulins, attached only to the corresponding anti-immunoglobulins This indicated that the method could be used for cell sorting. The attachment of cells to proteins was influnced, by the amount of bound protein, incubation time temperature and the degree of trypsinization. Most attached cells were viable for several days and when dying they detached. Detailed morphological and cytochemical analyses of the dynamics of attachment, and cytoplasmic spreading on the chemically well-defined surfaces were possible using the new method.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Rosenberg, M. D. 1960. Microexudates from cells grown in tissue culture. Biophys. J. 1: 137–159.
Brunk, U., J. L. F. Ericsson, J. Pontén, and B. Westermark. 1971. Specialization of cell surfaces in contact inhibited human glia-like cellsin vitro. Exp. Cell. Res. 67: 407–415.
Culp, L. A., and J. F. Buniel. 1976. Substrateattached serum and cell proteins in adhesion of mouse fibroblasts. J. Cell. Physiol. 88: 89–106.
Grinnell, F., and D. G. Hays. 1978. Induction of cell spreading by substratum-adsorbed ligands directed against the cell surface. Exp. Cell. Res. 116: 275–284.
Eagle, H., G. E. Foley, H. Koprowski, H. Lazarus, E. M. Levine, and R. A. Adams. 1970. Growth characteristics of virus transformed cells. J. Exp. Med. 131: 863–879.
Giard, D. J., S. A. Aaronson, G. J. Todaro, P. Arnstein, J. H. Kersey, H. Dosik, and W. P. Parks. 1973.In vitro cultivation of human tumors: Establishment, of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 51: 1417–1423.
Glimelius, B., B. Norling, B Westermark, and Å. Wasteson. 1979. A comparative study of human, normal and malignant glial, cells. J. Cell. Physiol. 98: 527–537.
Costachel, O., L. Fadei, and E. Badea. 1969. Tumour cell suspension culture on a nonadhesive substratum. Z. Krebsforsch. 72: 24–31.
Porath, J., R. Axén, and S. Ernbach. 1967. Chemical coupling of proteins to agarose. Nature 215: 1491–1492.
Lis, H., and N. Sharon. 1973. The biochemistry of plant lectins. Annu. Rev. Biochem. 42: 541–574.
Minowada, J., T. Ohnuma, and G. E. Moore. 1972. Rosette-forming human lymphoid cell lines. Establisyment and evidence for origin of thymus-derived lymphocytes. J. Natl. Cancer Inst. 49: 891–895.
Westermark, B., J. Pontén, and R. Hugosson. 1973. Determinants for the establishment of permanent tissue culture lines from human gliomas. Acta Pathol. Microbiol. Scand. Sect. A, 81: 791–805.
Pontén, J., B. Westermark, and R. Hugosson. 1960. Regulation of proliferation and movement of human glia-like cells in culture. Exp. Cell. Res. 58: 393–400.
Nilsson, K., and C. Sundström. 1974. Establishment and characteristics of two unique cell lines from patients with lymphosarcoma. Int. J. Cancer 13: 808–823.
Klein, E., G. Klein, J. S. Nadkarni, J. J. Nadkarni, H. Wigzell, and P. Clifford. 1968. Surface IgM-kappa specificity on a Burkitt lymphoma cellin vivo and in derived culture lines. Cancer Res. 28: 1300–1310.
Axén, R., and S. Ernbach. 1971. Chemical fíxation of enzymes to cyanogen halide activated polysaccharide carriers. Eur. J. Biochem. 18: 351–360.
Ham, R. G. 1963. An improved nutrient solution for diploid Chinese hamster and human cell lines. Exp. Cell. Res. 29: 515–526.
Kinzel, V., D. Kübler, J. Richards, and M. Stöhr. 1976. Lens culinaris lectin immobilized on sepharose. Binding and sugar-specific release of intact tissue culture cells. Science 192: 487–489.
Kolb, H. J., R. Renner, K. D. Hepp, L. Weiss, and O. H. Wieland. 1975. Reevaluation of sepharose-insulin as a tool for the study of insulin action. Proc. Natl. Acad. Sci. U.S.A. 72: 248–252.
Porath, J., J. C. Janson, and R. Låås. 1971. Agar derivatives for chromatography, electrophoresis and gel-bound enzymes. Desulphated and reduced cross-linked agar and agarose in spherical bead form. J. Chromatogr. 60: 167–177.
Sundberg, L., and J. Porath 1974. Preparation of adsorbents for biospecific affinity chromatography. Attachment of group-containing ligands to insoluble polymers by means of bifunctional oxiranes. J. Chromatogr. 90: 87–98.
Vretblad, P., and R. Axén. 1973. The use of isocyanides for the immobilization of biological molecules. Acta Chem. Scand. 27: 2769–2780.
Maroudas, N. G. 1973. Chemical and mechanical requirements for fibroblast adhesion. Nature 244: 353–354.
Giaever, I., and E. Ward. 1978. Cell adhesion to substrates containing adsorbed or attached IgG. Proc. Natl. Acad. Sci. U.S.A. 75: 1366–1368.
Alexander, E., and P. Henkart. 1976. The adherence of human Fc receptor-bearing lymphocytes to antigen-antibody complexes. II. Morphologic alterations induced by the substrate. J. Exp. Med. 143: 329–347.
Author information
Authors and Affiliations
Additional information
The work has been supported financially by the Swedish Cancer Society and the Swedish Natural Science Research Council.
Box 531.
Box 533.
Rights and permissions
About this article
Cite this article
Carlsson, J., Gabel, D., Larsson, E. et al. Protein-coated agarose surfaces for attachment of cells. In Vitro 15, 844–850 (1979). https://doi.org/10.1007/BF02618038
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02618038