All‐in‐one microfluidic design to integrate vascularized tumor spheroid into high‐throughput platform

Abstract

The development of a scalable and highly reproducible in vitro tumor microenvironment (TME) platform still sheds light on new insights into cancer metastasis mechanisms and anticancer therapeutic strategies. Here, we present an all-in-one injection molded plastic array three-dimensional culture platform (All-in-One-IMPACT) that integrates vascularized tumor spheroids for highly reproducible, high-throughput experimentation. This device allows the formation of self-assembled cell spheroids on a chip by applying the hanging drop method to the cell culture channel. Then, when the hydrogel containing endothelial cells and fibroblasts is injected, the spheroid inside the droplet can be patterned together in three dimensions along the culture channel. In just two steps above, we can build a vascularized TME within a defined area. This process does not require specialized user skill and minimizes error-inducing steps, enabling both reproducibility and high throughput of the experiment. We have successfully demonstrated the process, from spheroid formation to tumor vascularization, using patient-derived cancer cells (PDCs) as well as various cancer cell lines. Furthermore, we performed combination therapies with Taxol (paclitaxel) and Avastin (bevacizumab), which are used in standard care for metastatic cancer. The All-in-One IMPACT is a powerful tool for establishing various anticancer treatment strategies through the development of a complex TME for use in high-throughput experiments.all-in-one, high-throughput experimentation, microfluidics, organ-on-a-chip, patient-derived cancer cells (PDCs), tumor microenvironment (TME), vascularized tumor spheroid

Recent advances in vascularized tumor-on-a-chip

“…For in vitro vasculature formation, generally seeded at a density between 5 to 10 million cells/mL, together with supporting cells. Fibroblasts are the most widely used cell stromal cells, although other cell types have also been used depending on the microenvironment to be mimicked ( 13 – 18 ). The main reason for incorporating stromal cells is to support the formed microvasculature, which will otherwise regress after initial formation in the EC monoculture ( 19 ).…”

“…Kim et al. ( 18 ) recently developed an all-in-one IMPACT device, where spheroid formation and vascularization can be performed in the same device ( Figure 3A ). By injecting a small volume of cell suspension, the droplet was maintained by surface tension.…”

One-step achievement of tumor spheroid-induced angiogenesis in a high-throughput microfluidic platform: one-step tumor angiogenesis platform

“…The platform has the format of a 96-well plate and the size of a slide glass (1 inch 3 inches). The platform has 8 identical wells, each of which consists of 2 parts: a spheroid formation area and a cell culture region [28]. Both parts are designed to pattern a liquid for spheroid formation and reconstruction of the 3D microenvironment.…”

“…vascular networks, and performed experiments using 4 concentrations (1.25 mg/mL, 2.5 mg/mL, 5 mg/mL, and 10 mg/mL) of fibrin gel and control (2.5 mg/mL without tumor spheroids) to construct angiogenesis models. For tumor spheroids, we used gastric cancer PDCs that were used in the previous study [28]. Through the sample images that were obtained from confocal microscopy, we observed that tumor spheroid angiogenesis in 2.5 mg/mL fibrin gel showed the most sprouted vascular network compared to other concentrations and the control group (Fig.…”

Microfluidic Droplet-Assisted Fabrication of Vessel-Supported Tumors for Preclinical Drug Discovery

“…With the continuous development of microfabrication technology, microfluidic tumor–vessel chips not only help to precisely arrange the spatial distribution of cellular components in vitro but can also apply biomechanical and biochemical stimuli, including blood flow and growth factor gradient, thereby reproducing the interaction between the heterotypic tumor and TME with high fidelity. − For example, Haase et al reported an on-chip 3D-vascularized tumor model for drug examination in a TME within a large bed of perfusable vasculature. , They demonstrated that the dose-dependent effect of anticancer drugs on tumor activity was significantly affected by the form of vascular infusion administration. Meanwhile, great strides have also been made in the field of biomaterials. , As a representative biomaterial, GelMA has been widely used in the regenerative medicine field for 3D cell culture. , Various organ models have been successfully established in GelMA, which demonstrate its good biocompatibility. ,, Antunes et al reported an in-air production of tumor spheroid-laden GelMA microgel to expedite drug screening, where GelMA was involved as tumor–ECM .…”