Abstract
In vitro cell culture is traditionally performed within two-dimensional (2D) environments, providing a quick and cheap way to study cell properties in a laboratory. However, 2D systems differ from the in vivo environment and may not mimic the physiological cell behavior realistically. For instance, 2D culture models are thought to induce cancer stem cells (CSCs) differentiation, a rare cancer cell subpopulation responsible for tumor initiation and relapse. This fact hinders the development of therapeutic strategies for tumors with a high relapse percentage, such as triple negative breast cancer (TNBC). Thus, three-dimensional (3D) scaffolds have emerged as an attractive alternative to monolayer culture, simulating the extracellular matrix structure and maintaining the differentiation state of cells. In this work, scaffolds were fabricated through electrospinning different poly(ε-caprolactone)-acetone solutions. Poly(ε-caprolactone) (PCL) meshes were seeded with triple negative breast cancer (TNBC) cells and 15% PCL scaffolds displayed significantly (p < 0.05) higher cell proliferation and elongation than the other culture systems. Moreover, cells cultured on PCL scaffolds exhibited higher mammosphere forming capacity and aldehyde dehydrogenase activity than 2D-cultured cells, indicating a breast CSCs enrichment. These results prove the powerful capability of electrospinning technology in terms of poly(ε-caprolactone) nanofibers fabrication. In addition, this study has demonstrated that electrospun 15% PCL scaffolds are suitable tools to culture breast cancer cells in a more physiological way and to expand the niche of breast CSCs. In conclusion, three-dimensional cell culture using PCL scaffolds could be useful to study cancer stem cell behavior and may also trigger the development of new specific targets against such malignant subpopulation.
Highlights
In vitro cell culture represents a crucial tool to study cell behavior outside the organism.Most cell cultures are performed with a two-dimensional (2D) environment providing cheap and easy cell maintenance
To study microscopic scaffold architecture, both specimens were imaged by Scanning Electron Microscopy (SEM; Table 1)
Architecture, bothcharacterization specimens were imaged by Scanning and 15%Electron electrospun poly(SEM; (ε-caprolactone)
Summary
In vitro cell culture represents a crucial tool to study cell behavior outside the organism. Most cell cultures are performed with a two-dimensional (2D) environment providing cheap and easy cell maintenance. A flat plastic surface is treated, obtaining adherent features to enable cell adhesion and proliferation. These cells can only grow forming a monolayer, establishing interactions with surface and contiguous cells. Cells adopt a flattened morphology, which results in a modified membrane receptor polarity and cytoskeleton architecture.
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