Homogenization of Tissue Culture Cells in Suspension

For this procedure, the cells are first swollen in a buffered hypo-osmotic medium containing potassium chloride, magnesium acetate and dithiothreitol to render the plasma membrane more susceptible to subsequent homogenization. After pelleting the cells, most of the ‘swelling’ medium is removed; the cells are resuspended in the residual medium and homogenized by Dounce homogenization.

Keywords: homogenization; tissue culture; mammalial cells; monolayer cells; suspension cells

 References
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    Goldberg DE and Kornfeld S (1983) Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation. Journal of Biological Chemistry 258: 3159–3165.
    book Graham JM (1993) "Isolation of membranes from tissue culture cells". In: Graham JM and Higgins JA (eds) Methods in Molecular Biology, 19, pp.97–108. Totowa, NJ: Humana Press.
    book Graham JM (1997) "Homogenization of tissues and cells". In: Graham JM and Rickwood D (eds) Subcellular Fractionation – a Practical Approach, pp. 1–29. Oxford: Oxford University Press
    Marsh M, Schmid S, Kern H et al. (1987) Rapid analytical and preparative isolation of functional endosomes by free flow electrophoresis. Journal of Cell Biology 104: 875–886.
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Graham, John M(Mar 2004) Homogenization of Tissue Culture Cells in Suspension. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003803]