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  • Optimized Conditions for the Delivery of Small Membrane Impermeable Compounds into Human Cells Using Hypotonic Shift

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    Author(s)
    Stephens, Alexandre
    Day, Christopher
    Tiralongo, Joe
    Griffith University Author(s)
    Day, Christopher J.
    Stephens, Alexandre
    Tiralongo, Joe
    Year published
    2012
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    Abstract
    Cell-based assays represent a major end point of high throughput screening (HTS) but a key limitation of such assays is the potentially poor membrane permeability of test compounds. In this study, we optimized the conditions for the deliv-ery of the membrane impermeable compound 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) into human cells using hypotonic shift; a method that can promote the uptake of molecules from the extracellular fluid into cell cy-toplasm via endocytosis. We showed that uptake of HPTS by cells was a function of hypotonic buffer osmolarity and that delivery was highly efficient with almost ...
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    Cell-based assays represent a major end point of high throughput screening (HTS) but a key limitation of such assays is the potentially poor membrane permeability of test compounds. In this study, we optimized the conditions for the deliv-ery of the membrane impermeable compound 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) into human cells using hypotonic shift; a method that can promote the uptake of molecules from the extracellular fluid into cell cy-toplasm via endocytosis. We showed that uptake of HPTS by cells was a function of hypotonic buffer osmolarity and that delivery was highly efficient with almost 100% of cells displaying uptake. Delivery of HPTS was equally effective at 25àand 37ì with delivery of compound proportional to incubation time and concentration of HPTS within the loading medium. The experimental conditions identified in this study could be applied to HTS drug discovery studies providing an effective method of delivering small membrane impermeable compounds into cells.
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    Journal Title
    CellBio
    Volume
    1
    DOI
    https://doi.org/10.4236/cellbio.2012.12005
    Copyright Statement
    © 2012 Scientific Research Publishing. This is the author-manuscript version of this paper. Reproduced in accordance with the copyright policy of the publisher. Please refer to the journal website for access to the definitive, published version.
    Subject
    Receptors and Membrane Biology
    Analytical Biochemistry
    Publication URI
    http://hdl.handle.net/10072/49845
    Collection
    • Journal articles

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