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Metabolic cooperation studied by a quantitative enzyme assay of single cells
Authors:MP Uitendaal  TL Oei  CHMM de Bruyn  P Hösli
Institution:1. Department of Human Cenetics, Faculty of Medicine University of Nijmegen, Nijmegen, The Netherlands.;2. Department of Molecular Biology, Institut Pasteur, 25, Rue du Dr. Roux, Paris 15e, France.
Abstract:A new method making use of a radiochemical enzyme assay at the single cell level is presented to investigate metabolic cooperation, a widely studied form of cellular communication. In this case metabolic cooperation between normal human fibroblasts and fibroblasts derived from a patient deficient for the enzyme hypoxanthine-guanine phosphoribosyl transferase has been studied.A mixture of an equal number of both cell types was cultured in close physical contact and after trypsinisation, replating and culturing the cells for several hours in a high dilution, quantitative enzyme measurements with individual cells isolated from the mixture were carried out. From the distribution curve of the enzyme activities of the individual cells the conclusion could be drawn that a macromolecule, either the enzyme itself or DNA or mRNA, coding for that enzyme, is transferred from normal to mutant cells.
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