Configurable 2D and 3D spheroid tissue cultures on bioengineered surfaces with acquisition of epithelial–mesenchymal transition characteristics

Abstract

Both two-dimensional (2D) and three-dimensional (3D) biomaterial-based culture platforms that are capable of mimicking the in vivo microenvironment to recapitulate the physiological conditions are vital tools in a wide range of cellular and clinical research. Here we report tissue cultures in a microfluidic chip that allows deterministic patterning of cells in 2D/3D. The chip contains a cell-supporting membrane bioengineered to attain either 2D or 3D cell patterns by selectable deposition of extracellular matrix molecules. Results show a cell-trapping rate as high as 97% in our microchip. Tuning of the surface enables not only highly controlled geometry of the monolayer (2D) cell mass but also 3D culture of uniformly sized multicellular spheroids. The 3D spheroid culture of human epithelial ovarian cancer cells in the microfluidic chip resulted in acquisition of mesenchymal traits-increased expressions of N-cadherin, vimentin and fibronectin-and lowered expression of epithelial marker (CD326/epithelial cell adhesion molecule) compared with that in traditional 2D cultures, which is indicative of epithelial-mesenchymal transition in the spheres. In conclusion, these results offer new opportunities to achieve active control of 2D cellular patterns and 3D multicellular spheroids on demand, and may be amenable toward the study of the metastatic processes by in vitro modeling.

Assessing nanoparticle toxicity in cell-based assays: influence of cell culture parameters and optimized models for bridging the in vitro–in vivo gap

“…In turn, the enhanced communication influences a number of important cellular functions such as migration, invasion, proliferation, apoptosis and differentiation 209 . It was, for example, shown by Kuo et al that the epithelialmesenchymal-transition, which is a hallmark in the metastatic process, is promoted for several cancer cell lines when they are cultured as spheres 210 . Thus, it can be concluded that the cultivation conditions have a major influence on the cellular phenotype and function and that cellular responses following NP exposure are therefore likely to be drastically altered in a 3D model when compared to a conventional monolayer 207 .…”

Inventions and Innovations in Preclinical Platforms for Cancer Research

“…Kuo et al reported a size standard deviation of 104% for on dish liquid overlay and 13% for on-chip spheroid diameters [129]. This illustrated that the spheroid size was much more uniform in the microfluidic approach in comparison with that in the liquid overlay technique.…”

Observation of “wired” cell communication over 10-μm and 20-μm poly(dimethylsiloxane) barriers in tetracycline inducible expression systems

“…Afterwards, PBS and culture medium were introduced to wash away unbound collagen. 100 ll of HEK 293T cells ($1.5 Â 10 3 cells) in culture medium were perfused into the top channel by tuning the solution level height as described, 8 and the device was placed in a 5% CO 2 incubator at 37 C overnight to enable cell attachment and spread on the membrane. Subsequently, Tet was introduced into the bottom channel at t ¼ 0 h, as on-device experiments.…”