Colorectal Adenocarcinoma Cell Culture in a Microfluidically Controlled Environment with a Static Molecular Gradient of Polyphenol

Abstract

To study the simultaneous effect of the molecular gradient of polyphenols (curcumin, trans-resveratrol, and wogonin) and biological factors released from tumor cells on apoptosis of adjacent cells, a novel microfluidic system was designed and manufactured. The small height/volume of microfluidic culture chambers and static conditions allowed for establishing the local microenvironment and maintaining undisturbed concentration profiles of naturally secreted from cells biochemical factors. In all trials, we observe that these conditions significantly affect cell viability by stimulating cell apoptosis at lower concentrations of polyphenols than in traditional multiwell cultures. The observed difference varied between 20.4–87.8% for curcumin, 11.0–37.5% for resveratrol, and 21.7–62.2% for wogonin. At low concentrations of polyphenols, the proapoptotic substances released from adjacent cells, like protein degradation products, significantly influence cell viability. The mean increase in cell mortality was 38.3% for microfluidic cultures. Our research has also confirmed that the gradient microsystem is useful in routine laboratory tests in the same way as a multiwell plate and may be treated as its replacement in the future. We elaborated the new repetitive procedures for cell culture and tests in static gradient conditions, which may become a gold standard of new drug investigations in the future.

The Lattice-Boltzmann Modeling of Microflows in a Cell Culture Microdevice for High-Throughput Drug Screening

“…The conclusions drawn from the simulation results are consistent with the observations made previously during the practical tests of the device [20]. In a further article, we intend to present an in-depth analysis of research results on the formation of active substance concentration profiles in CC and their stability over time.…”

“…We have analyzed the liquid flow hydrodynamics in the device's CC, performing numerical simulations of the LBM/CFD method for the same geometry and conditions as present in the device's standard operation protocols [20]. The simulation results were compared with the experimentally determined overall volumetric flow rates of the fluid in CC.…”

“…The main structural elements of the microsystem are three identical cell culture chambers with dimensions of 59/4.1/0.13 mm (L/W/H). A detailed discussion of chip design as well as analytical and cell culture procedures was presented in the previous publication [20].…”

“…The main structural elements of the microsystem are three identical cell culture chambers with dimensions of 59/4.1/0.13 mm (L/W/H). A detailed discussion of chip design as well as analytical and cell culture procedures was presented in the previous publication [20]. The device's simple operation protocols do not require specialized equipment (actuators, pumps, control valves) or costly automated equipment, such as a pipetting robot.…”

Analysis of Static Molecular Gradients in a High-Throughput Drug Screening Microfluidic Assay

“…Recently, we published a study where we introduced a three-chamber microfluidic device for cell culture in static gradients [27]. The device was used to analyze induced apoptosis of adenocarcinoma (LOVO) cells by three polyphenols: curcumin, trans-resveratrol, and wogonin.…”