Detachably assembled microfluidic device for perfusion culture and post‐culture analysis of a spheroid array

Abstract

Microfluidic devices permit perfusion culture of three-dimensional (3D) tissue, mimicking the flow of blood in vascularized 3D tissue in our body. Here, we report a microfluidic device composed of a two-part microfluidic chamber chip and multi-microwell array chip able to be disassembled at the culture endpoint. Within the microfluidic chamber, an array of 3D tissue aggregates (spheroids) can be formed and cultured under perfusion. Subsequently, detailed post-culture analysis of the spheroids collected from the disassembled device can be performed. This device facilitates uniform spheroid formation, growth analysis in a high-throughput format, controlled proliferation via perfusion flow rate, and post-culture analysis of spheroids. We used the device to culture spheroids of human hepatocellular carcinoma (HepG2) cells under two controlled perfusion flow rates. HepG2 spheroids exhibited greater cell growth at higher perfusion flow rates than at lower perfusion flow rates, and exhibited different metabolic activity and mRNA and protein expression under the different flow rate conditions. These results show the potential of perfusion culture to precisely control the culture environment in microfluidic devices. The construction of spheroid array chambers allows multiple culture conditions to be tested simultaneously, with potential applications in toxicity and drug screening.

Engineering Functional Pseudo-Islets of Defined Sizes from Primary Murine Cells Using PEG Microwell Devices

“…The use of microwell arrays has been exploited for use with a range of cell types [44][45][46][47], and has been recently utilized to create β-cell clusters [37,38,48].…”

Patterning hypoxic multicellular spheroids in a 3D matrix – a promising method for anti‐tumor drug screening

“…Patterning hypoxic multicellular spheroids in a 3D matrixa promising method for anti-tumor drug screening spheroids are heterogeneous cellular aggregates that are often characterized by multicellular components within a 3D extracellular matrix and hypoxic or necrotic regions. These multicellular tumor spheroids adopt similar phenotypes and respond to stimuli analogous to in vivo biological systems [14][15][16][17]. Thus, 3D spheroids are considered to be valid models for reproducing the properties of tumor microregions, intervascular regions and micrometastases [18,19].…”

Time‐dependent structural and functional characterization of subcutaneous human liver tissue

“…In addition, the hepatic organoids could be used as grafts for cell therapies for liver diseases. However, hepatocytes have shown to have particularly high metabolic activity, resulting in high frequencies of supply limitation of oxygen and nutrients inside the hepatic organoids (Anada, Fukuda, Sai, & Suzuki, ; De Bartolo et al, ; Sakai et al, ; Sakai & Nakazawa, ). Thus, the organoid size and/or supply system for oxygen and nutrients require successful design.…”