A micro cell culture analog (µCCA) with 3-D hydrogel culture of multiple cell lines to assess metabolism-dependent cytotoxicity of anti-cancer drugs

Abstract

A microfluidic device with 3-D hydrogel cell cultures has been developed to test the cytotoxicity of anti-cancer drugs while reproducing multi-organ interactions. In this device, a micro cell culture analog (microCCA), cells embedded in 3-D hydrogels are cultured in separate chambers representing the liver, tumor, and marrow, which are connected by channels mimicking blood flow. While the microfluidic network provides a platform for mimicking the pharmacokinetic and pharmacodynamic profiles of a drug in humans, the 3-D hydrogel provides a more physiologically realistic environment to mimic the tissue than monolayer culture. Colon cancer cells (HCT-116) and hepatoma cells (HepG2/C3A) were encapsulated in Matrigel and cultured in the tumor and the liver chamber in a microCCA, respectively. Myeloblasts (Kasumi-1) were encapsulated in alginate in the marrow chamber; a stiffer hydrogel was necessary to prevent cell migration out of the matrix. The cytotoxic effect of Tegafur, an oral prodrug of 5-fluorouracil (5-FU), on each cell line was tested using the microCCA with cell-embedded hydrogel. The comparison of experimental results using a 96-well microtiter plate and a microCCA demonstrated that the microCCA was able to reproduce the metabolism of Tegafur to 5-FU in the liver and consequent death of cells by 5-FU, while the cultures in a 96-well microtiter plate were unable to do so. The microCCA utilizing 3-D hydrogel cell cultures has potential as a platform for pharmacokinetic-based drug screening in a more physiologically realistic environment.

Reconfigurable microfluidic hanging drop network for multi-tissue interaction and analysis

“…4a). We realized a hepatic pro-drug activation experiment, which differs from other recent work 17 , in the used pro-drugs, cell types, device structure (cells in chambers with hydrogels) and microfluidic approach. The array consisted of four columns and six rows.…”

“…A next logical step is the development of fluidically interconnected networks of 3D tissue structures, potentially derived from different cell types, which paves the way to a new generation of model systems for complex multi-tissue studies with the aim to ultimately being able to mimic the behaviour of larger organ networks of the human body ('body-on-a-chip' research initiatives) [12][13][14][15][16][17][18][19][20][21] .…”

Chip-based liver equivalents for toxicity testing – organotypicalness versus cost-efficient high throughput

“…2b) has been improved regarding culture format from monolayer hepatocyte cultures to 3D hydrogel supported cultures. 55 It combines cultures of human hepatoma cells HepG2/C3A with colon cancer cells and myeloblasts in separate cell culture chambers. The system, at its selected scale, provides in vivo-like tissue mass ratios inside the culture compartments, a culture medium inflow split that is equivalent to the respective blood flow split in humans, and relevant residence times in the tissue compartments.…”

Modeling Life

“…Our initial devices used cell lines and 2D culture, and the ratio of free liquid to cells was higher than physiologic. For more recent devices 20 , 3D tissue constructs based on hydrogel entrapped cells have been used, and this technique facilitates design of compartments with more physiologic ratios of liquid to cells. The last issue of authentic physiological response of cells or tissues in a compartment is the most problematic.…”

“…Thus, using PBPK models and experimental results, the investigator should be able to obtain a deeper insight into possible underlying mechanisms. 19,20 Another advantage of the microCCA approach is the extrapolation of response of animals to humans. For example, rat tissue can be used in a microCCA and tested with a drug; the predicted results can then be confirmed with animal studies.…”