A microfluidic-based neurotoxin concentration gradient for the generation of an in vitro model of Parkinson’s disease

Abstract

In this study, we developed a miniaturized microfluidic-based high-throughput cell toxicity assay to create an in vitro model of Parkinson's disease ͑PD͒. In particular, we generated concentration gradients of 6-hydroxydopamine ͑6-OHDA͒ to trigger a process of neuronal apoptosis in pheochromocytoma PC12 neuronal cell line. PC12 cells were cultured in a microfluidic channel, and a concentration gradient of 6-OHDA was generated in the channel by using a back and forth movement of the fluid flow. Cellular apoptosis was then analyzed along the channel. The results indicate that at low concentrations of 6-OHDA along the gradient ͑i.e., approximately less than 260 M͒, the neuronal death in the channel was mainly induced by apoptosis, while at higher concentrations, 6-OHDA induced neuronal death mainly through necrosis. Thus, this concentration appears to be useful for creating an in vitro model of PD by inducing the highest level of apoptosis in PC12 cells. As microfluidic systems are advantageous in a range of properties such as throughput and lower use of reagents, they may provide a useful approach for generating in vitro models of disease for drug discovery applications.

From 3D cell culture to organs-on-chips

“…It also has been extended to study more complex cell behaviors, such as stem cell differentiation [15–17], axon guidance [18], subcellular propagation of biochemical signaling [19], and embryonic development [20, 21] in 2D culture, and to develop novel disease models (e.g. Parkinson’s disease; [22]).…”

Gezielte dreidimensionale Zellausrichtung und -elongation in artifiziellen Geweben

“…Dennoch bleibt der Nutzen von Oberflächenstrukturen oder von präformier-ten Kanälen zur gezielten Zellsteuerung auf Mikrometerniveau zunächst auf In-vitro-Studienmodelle beschränkt, wie beispielweise in der Diagnostik [17] oder der Medikamententestung [18]. Dies ist insbesondere darauf zurückzuführen, dass man die Zellen immer nur auf einer 2D-Ebene beeinflussen kann, und dieselben Verfahren nicht ohne weiteres auf eine komplexere Komposition, etwa in einem dreidimensionalen Gewebeverband übertragen werden können.…”

Multi-dimensional studies of synthetic genetic promoters enabled by microfluidic impact printing

“…17 Both methods permit concentration gradients to be maintained over an extended period of time, on the order of hours. 24 Recent introduction to droplet microfluidics provides an alternative approach to generate gradient, in which multi-component aqueous droplets can be physically compartmentalized from samples merged from different flow channels at different flow rates. Droplet microfluidics allow for high-throughput gradient generation and analysis with extremely low sample consumption (pico-liter to nano-liter reactant volumes), effectively enabling this technology to adapt to copious uses in biological assaying, including protein crystallization concentration screening 25 and enzyme inhibition assays.…”