Abstract
Tissues are complex three-dimensional structures in which cell behaviour is frequently guided by chemotactic signals. Although starvation and nutrient restriction induce many different chemotactic processes, the recreation of such conditions in vitro remains difficult when using standard cell culture equipment. Recently, microfluidic techniques have arisen as powerful tools to mimic such physiological conditions. In this context, microfluidic three-dimensional cell culture systems require precise control of cell/hydrogel location because samples need to be placed within a microchamber without obstruction of surrounding elements. In this article, SU-8 is studied as structural material for the fabrication of complex cell culture devices due to its good mechanical properties, low gas permeability and sensor integration capacity. In particular, this manuscript presents a SU-8 based microdevice designed to create “self-induced” medium starvation, based on the combination of nutrient restriction and natural cell metabolism. Results show a natural migratory response towards nutrient source, showing how cells adapt to their own microenvironment modifications. The presented results demonstrate the SU-8 potential for microdevice fabrication applied to cell culture.
Highlights
Living tissues are composed of embedded cells within a heterogeneous extracellular matrix, which plays a key role in three-dimensional cell migration and intercellular communication (Raines, 2000; Savino et al, 2004; Korpos et al, 2010; Franco and Muller, 2011)
The results presented in this paper validate SU-8 based microfluidic devices as a novel and robust tool for future three-dimensional self-induced cell migration assays and allow studying cell behavior under nutrient starvation
Microdevice Design In order to establish three-dimensional cell culture, microfluidic devices are usually designed with a central culture microchamber and lateral microchannels at both sides that are delimited with a series of pillars (Figure 1)
Summary
Living tissues are composed of embedded cells within a heterogeneous extracellular matrix, which plays a key role in three-dimensional cell migration and intercellular communication (Raines, 2000; Savino et al, 2004; Korpos et al, 2010; Franco and Muller, 2011). Microfabrication and microfluidic technologies have arisen as interesting alternatives for creating high-performance cell culture systems (El-Ali et al, 2006; Kim et al, 2010; Lesher-Perez et al, 2013) Such devices possess a culture microchamber housing a hydrogel and lateral microchannels delimited by a series of pillars (Figure 1). The results presented in this paper validate SU-8 based microfluidic devices as a novel and robust tool for future three-dimensional self-induced cell migration assays and allow studying cell behavior under nutrient starvation. In this article, this methodology is validated in the context of bone tissue repair, since blood vessels are absent in bone scaffolds and nutrient support is compromised. The described methodology could be applied in many different biological scenarios
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