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    Cell-free DNA in a three-dimensional spheroid cell culture model: a preliminary study

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    Date
    2017
    Author
    Aucamp, Janine
    Calitz, Carlemi
    Bronkhorst, Abel J.
    Hamman, Sias
    Gouws, Chrisna
    Pretorius, Piet J.
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    Abstract
    Background Investigating the biological functions of cell-free DNA (cfDNA) is limited by the interference of vast numbers of putative sources and causes of DNA release into circulation. Utilization of three-dimensional (3D) spheroid cell cultures, models with characteristics closer to the in vivo state, may be of significant benefit for cfDNA research. Methods CfDNA was isolated from the growth medium of C3A spheroid cultures in rotating bioreactors during both normal growth and treatment with acetaminophen. Spheroid growth was monitored via planimetry, lactate dehydrogenase activity and glucose consumption and was related to isolated cfDNA characteristics. Results Changes in spheroid growth and stability were effectively mirrored by cfDNA characteristics. CfDNA characteristics correlated with that of previous two-dimensional (2D) cell culture and human plasma research. Conclusions 3D spheroid cultures can serve as effective, simplified in vivo-simulating “closed-circuit” models since putative sources of cfDNA are limited to only the targeted cells. In addition, cfDNA can also serve as an alternative or auxiliary marker for tracking spheroid growth, development and culture stability. Biological significance 3D cell cultures can be used to translate “closed-circuit” in vitro model research into data that is relevant for in vivo studies and clinical applications. In turn, the utilization of cfDNA during 3D culture research can optimize sample collection without affecting the stability of the growth environment. Combining 3D culture and cfDNA research could, therefore, optimize both research fields
    URI
    http://hdl.handle.net/10394/25314
    https://www.sciencedirect.com/science/article/pii/S1357272517301528
    https://doi.org/10.1016/j.biocel.2017.06.014
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    • Faculty of Health Sciences [2404]
    • Faculty of Natural and Agricultural Sciences [4855]

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