A 3D printed microfluidic perfusion device for multicellular spheroid cultures

Abstract

The advent of 3D printing technologies promises to make microfluidic organ-on-chip technologies more accessible for the biological research community. To date, hydrogel-encapsulated cells have been successfully incorporated into 3D printed microfluidic devices. However, there is currently no 3D printed microfluidic device that can support multicellular spheroid culture, which facilitates extensive cell–cell contacts important for recapitulating many multicellular functional biological structures. Here, we report a first instance of fabricating a 3D printed microfluidic cell culture device capable of directly immobilizing and maintaining the viability and functionality of 3D multicellular spheroids. We evaluated the feasibility of two common 3D printing technologies i.e. stereolithography (SLA) and PolyJet printing, and found that SLA could prototype a device comprising of cell immobilizing micro-structures that were housed within a microfluidic network with higher fidelity. We have also implemented a pump-free perfusion system, relying on gravity-driven flow to perform medium perfusion in order to reduce the complexity and footprint of the device setup, thereby improving its adaptability into a standard biological laboratory. Finally, we demonstrated the biological performance of the 3D printed device by performing pump-free perfusion cultures of patient-derived parental and metastatic oral squamous cell carcinoma tumor and liver cell (HepG2) spheroids with good cell viability and functionality. This paper presents a proof-of-concept in simplifying and integrating the prototyping and operation of a microfluidic spheroid culture device, which will facilitate its applications in various drug efficacy, metabolism and toxicity studies.

A Review of Current Methods in Microfluidic Device Fabrication and Future Commercialization Prospects

“…The volume under the glass coverslip was filled with the GelMa resin, and it was synchronized with a moving platform and changing images to cure a structure that encapsulated the cells between the cover-slip and structure ( Figure 5D). A multicellular device was fabricated by Ye Ong et al [62] for the study of spheroid cultures. The device was fabricated using both SL and Polyjet printing using commercial printers.…”

“…The issue with this technology is that the MJM machines tend to be expensive, and most resins are proprietary [42]. Also, MJM can have difficulty producing features in the submillimeter range [62].…”

Microfluidic Model for Evaluation of Immune Checkpoint Inhibitors in Human Tumors

“…The TAP device utilizes fresh tumor biopsy samples, permitting evaluation within 24 h of resection from a patient. More typical tumor recapitulation platforms make use of organoid and spheroid models (i.e., aggregations of cells grown in vitro) which have shown promise in cancer research applications and early pharmaceutical evaluations . However, as predictive tools for real‐time and personalized treatment, the utility of these synthetic models is limited.…”

3D human bone marrow stromal and endothelial cell spheres promote bone healing in an osteogenic niche

“…Over the last 20 years, 3-dimensional (3D) spheroid models have been recognized by cell biologists and tissue engineers as an important tool to study cell-cell interaction together with the extracellular matrix created in these spheroids (9)(10)(11)(12)(13)(14). The cell-to-cell contact within pellet cultures is dictated by signaling proteins, including integrins to stimulate the secretion of extracellular matrix that can aid the natural growth and movement of cells (9,(15)(16)(17).…”