Cell-free DNA in a three-dimensional spheroid cell culture model: a preliminary study
Date
2017Author
Aucamp, Janine
Calitz, Carlemi
Bronkhorst, Abel J.
Hamman, Sias
Gouws, Chrisna
Pretorius, Piet J.
Metadata
Show full item recordAbstract
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/10394/25314https://www.sciencedirect.com/science/article/pii/S1357272517301528
https://doi.org/10.1016/j.biocel.2017.06.014