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Abstract
In vitro cell based models have been invaluable tools for studying cell behaviour and for investigating drug disposition, toxicity and potential adverse effects of administered drugs. Within this drug discovery pipeline, the ability to assess and prioritise candidate compounds as soon as possible offers a distinct advantage. However, the ability to apply this approach to a cell culture study is limited by the need to provide an accurate, in vitro-like, microenvironment in conjunction with a low cost and high-throughput screening (HTS) methodology. Although the geometry and/or alignment of cells has been reported to have a profound influence on cell growth and differentiation, only a handful of studies have directly compared the growth of a single cell line on different shaped multiwell plates the most commonly used substrate for HTS, in vitro, studies. Herein, the impact of various surface geometries (flat, round and v-shaped 96 well plates), as well as fixed volume growth media and fixed growth surface area have been investigated on the characteristics of three commonly used human cell lines in biopharmaceutical research and development, namely ARPE-19 (retinal epithelial), A549 (alveolar epithelial) and Malme-3M (dermal fibroblastic) cells. The effect of the surface curvature on cells was characterised using a combination of a metabolic activity assay (CellTiter AQ/MTS), LDH release profiles (CytoTox ONE) and absolute cell counts (Guava ViaCount), respectively. In addition, cell differentiation and expression of specific marker proteins were determined using flow cytometry. These in vitro results confirmed that surface topography had a significant effect (p < 0.05) on cell activity and morphology. However, although specific marker proteins were expressed on day 1 and 5 of the experiment, no significant differences were seen between the different plate geometries (p < 0.05) at the later time point. Accordingly, these results highlight the impact of substrate geometry on the culture of a cell line and the influence it has on the cells’ correct growth and differentiation characteristics. As such, these results provide important implications in many aspects of cell biology the development of a HTS, in vitro, cell based systems to further investigate different aspects of toxicity testing and drug delivery.
Highlights
In vitro cell culture models provide simple, fast and cost-effective tools for biological cell research and help to minimise the exploitation of animal testing [1]
Considerations are required to address the balance between using more complete in vivo-like models and the ability to generate a high-throughput screening (HTS) methodology to expedite a typical drug discovery pipeline process
Consideration to ensure that both fixed surface area (FSA) and fixed volume (FV) during culture needs to be maintained within the appropriate experimental set-up
Summary
In vitro cell culture models provide simple, fast and cost-effective tools for biological cell research and help to minimise the exploitation of animal testing [1]. Traditional long-standing two-dimensional (2D) cell culture models are based on the growth of specific cells on flat and rigid culture substrates/scaffolds within a controlled laboratory environment. These cells are themselves classified into three distinct groups namely, (i) adherent cells which must attach to a solid substrate during culture, (ii) suspension-based cells which are cultured as “floating units” within the culture medium [2], and (iii) cells that exhibit a mixed adherent-suspension characteristic. The biggest disadvantage of such culture systems is that it does not fully replicate the microenvironment experienced in vivo where cells grow within a complex three-dimensional (3D) matrix and, as the 3D structure impacts biological processes from the molecular level (i.e. gene and protein synthesis, and biomolecular gradients) [3] to the proliferation, differentiation and apoptotic nature of the cells, consideration of this key factor must be sought [4]
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