Gravity-oriented microfluidic device for uniform and massive cell spheroid formation

Abstract

We propose a simple method for forming massive and uniform three-dimensional (3-D) cell spheroids in a multi-level structured microfluidic device by gravitational force. The concept of orienting the device vertically has allowed spheroid formation, long-term perfusion, and retrieval of the cultured spheroids by user-friendly standard pipetting. We have successfully formed, perfused, and retrieved uniform, size-controllable, well-conditioned spheroids of human embryonic kidney 293 cells (HEK 293) in the gravity-oriented microfluidic device. We expect the proposed method will be a useful tool to study in-vitro 3-D cell models for the proliferation, differentiation, and metabolism of embryoid bodies or tumours.

A microfluidic device for uniform-sized cell spheroids formation, culture, harvesting and flow cytometry analysis

“…25 Formation of cell spheroids using gravity oriented microfluidic device was also introduced. 26 However, current spheroid culture devices possess some drawbacks that retard their practical use. The multilayer device with semi-transparent membranes suffers from problems of high fidelity imaging and real time monitoring.…”

“…Recently, researchers have developed various spheroid formation and culture devices based on microfluidics techniques. [18][19][20][21][22][23][24][25][26][27][28][29][30][31][32] A multilayer microfluidic device with a porous membrane was employed to achieve both the spheroid formation and in-situ culture. 24 A microfluidic array platform containing concave microwells and flat cell culture chambers for EB formation and its culture was also developed.…”

“…24 In addition, the spheroids cannot be easily harvested from the devices due to their channel designs without additional instrumentation. 26,29 The conventional analysis techniques include fluorescence staining using the antibody tagged fluorophores, but most of the microfluidic devices cannot form and culture a large number of cell spheroids with uniform size and harvest them out for further conventional analysis, such as flow cytometry or western blot.…”

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].…”

“…Cell spheroids have been used as 3D cell cultures for mesenchymal stem cells (MSCs) [18], liver tissue [19], cardiac muscle [20], human embryonic kidney cells [21] and so forth. Embryonic stem cells, neural stem cells, pancreatic cells and hepatocytes also need to be cultured in 3D configurations to induce differentiation and express their own metabolism and proliferation rate similar to the in vivo conditions.…”