IMPROVED DIFFUSION CHAMBER CULTURES FOR CYTOKINETIC ANALYSIS OF ANTIBODY RESPONSE

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  • 1 January 1966
    • journal article
    • research article
    • Vol. 11  (5) , 427-+
Abstract
Diffusion chambers (3 x 10 mm) constructed with 0[center dot]l [mu] porosity filters, but not with 0-45 [mu] or greater porosity filters, were found to be consistently cell impermeable, with use of acryloid as the glueing agent. The filters permit free diffusion of 19S and 7S antibodies into ''empty'' chambers in vivo and in vitro. Pronase treatment of the chamber dissolves the clot and frees cells attached to the inner surfaces. This permits almost complete recovery of the chamber culture cells. Chamber cultures can be readily transferred from one host to another and kept in vitro at room temperature for at least 6 hr. without any loss of activity. In vivo diffusion problems arise after 1 month of culture, most probably due to excessive growth of peritoneal cells on the outer surface of the filters; this limitation can be overcome by serial in vivo transfer of the chamber and wiping the outer surface at the time of transfer. The diffusion chamber culture method as described here fulfills all the prerequisites of an assay system with which one can perform precise cytokinetic analysis of antibody response.