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Protein-coated agarose surfaces for attachment of cells
Authors:J. Carlsson  D. Gabel  E. Larsson  J. Pontén  B. Westermark
Affiliation:(1) Department of Physical Biology, The Gustaf Werner Institute, University of Uppsala, S-751 21 Uppsala, Sweden;(2) The Institute of Biochemistry, University of Uppsala, S-751 21 Uppsala, Sweden;(3) The Wallenberg Laboratory, University of Uppsala, S-751 21 Uppsala, Sweden;(4) Present address: Department of Chemistry, University of Bremen, Bremen, Federal Republic of Germany
Abstract: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. The work has been supported financially by the Swedish Cancer Society and the Swedish Natural Science Research Council. Box 531. Box 533.
Keywords:cell attachment  cell culture  Con-A  immunoglobulins  membrane receptors  trypsinization
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