A microfluidic-based platform for tumour spheroid culture, monitoring and drug screening

Abstract

The development of novel cellular models that can replace animals in preclinical trials of drug candidates is one of the major goals of cell engineering. Current in vitro screening methods hardly correspond with the in vivo situation, whereas there is a lack of assays for more accurate cell culture models. Therefore, development of automated assays for 3D cell culture models is urgently required. In this work, we present a SpheroChip system: a microfluidic-based platform for long-term 3D cell culture and analysis. The system is compatible with commercially available microplate readers and provides continuous, in situ monitoring of tumour spheroids cultured on a chip. The microfluidic chip consists of cell culture microchambers and hemispherical microwells connected with a concentration gradient generator. HT-29 and Hep-G2 cells were successfully cultured as tumour spheroids in the SpheroChip, and metabolic activity of cells was monitored for up to two weeks by in situ fluorimetric measurements. Cellular response to an anticancer drug was observed using the SpheroChip. The experimental setup provided the unique possibility of observing dynamic changes in metabolic activity of one culture during sequencing days after drug dosage. According to this new approach, unknown phenomena of cellular response to the anticancer drug were observed, such as increase of metabolic activity shortly after drug dosage. Moreover, the influence of a second dose of a drug was evaluated. The SpheroChip system can be used by researchers working on drug screening, evaluation of anticancer procedures and chemoresistance phenomena.

Advances in multicellular spheroids formation

“…The micro-moulding method has been explored to generate MSCs by fabricating complex structures using the lithography technique [109,110]. It has been documented that this micro moulding method can significantly reduce consumption of reagents, create desired concentration gradients of growth/signal factors and nutrients, allow high-density culture at a high cell-to-fluid volume ratio and decrease shear stress under the laminar condition in micro-scale structures [111].…”

Inventions and Innovations in Preclinical Platforms for Cancer Research

“…Microwells [8,25,[87][88][89] and U-shaped microstructures [21,64,[90][91][92][93] have been designed for spheroid formation and culture in microfluidic platforms. These structures facilitate short-term [23,94], controllable and uniform diameter [22,95] and compact spheroid generation [32,92].…”

Multiphase microfluidic synthesis of micro- and nanostructures for pharmaceutical applications

“…Kwapiszewska et al developed the SpheroChip, consisting of disposable microfluidic chips for cell culture and a positioning plate, to culture human cell lines (i.e., HT-29 colon cancer cells and Hep-G2 hepatocytes). After the injection of anticancer drug 5-fluorouracil (5-FU), the dynamic changes on the metabolic activity of the cells in response to the drug could be monitored and analyzed continuously (Kwapiszewska et al, 2014). Ruppen et al observed a higher chemoresistance in primary co-culture 33 spheroids (i.e., primary lung cancer epithelial cells and primary pericytes) than in primary monoculture spheroids in a Lung-on-a-Chip model, which could serve as a more chemosensitive tumor model (Ruppen et al, 2015).…”

“…Skin melanoma -Association between cancer and immune system (Businaro et al, 2013) Colon cancer Anticancer drugs (e.g., 5-fluorouracil) Anticancer efficacy; drug screening (Kwapiszewska et al, 2014) Breast cancer -Invasion (Trietsch et al, 2013) Photosensitizer (e.g., 5-aminolevulinic acid) and gold nanoparticles Photodynamic therapy evaluation (Yang et al, 2015) Lung cancer Anticancer drugs (e.g., gefitinib and masitinib) Anticancer efficacy; drug screening (Aref et al, 2013) Cisplatin Chemosensitivity evaluation (Ruppen et al, 2015) Brain glioma Anticancer drugs (e.g., vincristine and bleomycin)…”