Abstract
Spheroid, a 3D aggregate of tumor cells in a spherical shape, has overcome the limitations of conventional 3D cell models to accurately mimic the in-vivo environment of a human body. The spheroids are cultured with other primary cells and embedded in collagen drops using hang drop plates and low-attachment well plates to construct a spheroid–hydrogel model that better mimics the cell–cell and cell–extracellular matrix (ECM) interactions. However, the conventional methods of culturing and embedding spheroids into ECM have several shortcomings. The procedure of transferring a single spheroid at a time by manual pipetting results in well-to-well variation and even loss or damage of the spheroid. Based on the previously introduced droplet contact-based spheroid transfer technique, we present a poly(dimethylsiloxane) and resin-based drop array chip and a pillar array chip with alignment stoppers, which enhances the alignment between the chips for uniform placement of spheroids. This method allows the facile and stable transfer of the spheroid array and even eliminates the need for a stereomicroscope while handling the cell models. The novel platform demonstrates a homogeneous and time-efficient construction and diverse analysis of an array of fibroblast-associated glioblastoma multiforme spheroids that are embedded in collagen.
Highlights
For basic and applied research of disease, efficient development of new drugs, innovative development of precision medicine, and reduction of animal experimentation, it is necessary to construct an in vitro cell model that precisely imitates the in vivo tumor environment
Type, and organization of the protein-based hydrogel, the tumor spheroid-embedded hydrogel models serve as a useful platform for investigating invasive and metastatic characteristics, effects of drug treatment, and other biochemical cues that regulate tumor cell behavior [18]
The drop array chip (DAC) consists of a PDMS-based concave well array assembled on the center of the plasCLEAR-based plate with alignment holes
Summary
For basic and applied research of disease, efficient development of new drugs, innovative development of precision medicine, and reduction of animal experimentation, it is necessary to construct an in vitro cell model that precisely imitates the in vivo tumor environment. The conventional two-dimensional (2D) cell models significantly differ from the in vivo environment, producing cellular behavior dissimilar to that of natural conditions. This is because the cell-to-cell and cell-to-extracellular matrix (ECM) interactions within the actual body, as well as the expression of genes, are not properly achieved [1,2,3]. Type, and organization of the protein-based hydrogel, the tumor spheroid-embedded hydrogel models serve as a useful platform for investigating invasive and metastatic characteristics, effects of drug treatment, and other biochemical cues that regulate tumor cell behavior [18]
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