High-throughput fabrication of cell spheroids with 3D acoustic assembly devices

Abstract

Acoustic cell assembly devices are applied in cell spheroid fabrication attributed to their rapid, label-free and low-cell damage production of size-uniform spheroids. However, the spheroids yield and production efficiency are still insufficient to meet the requirements of several biomedical applications, especially those that require large quantities of cell spheroids, such as high-throughput screening, macro-scale tissue fabrication, and tissue repair. Here, we developed a novel 3D acoustic cell assembly device combined with a gelatin methacrylamide (GelMA) hydrogels for the high-throughput fabrication of cell spheroids. The acoustic device employs three orthogonal piezoelectric transducers that can generate three orthogonal standing bulk acoustic waves to create a 3D dot-array (25 × 25 × 22) of levitated acoustic nodes, enabling large-scale fabrication of cell aggregates (>13,000 per operation). The GelMA hydrogel serves as a supporting scaffold to preserve the structure of cell aggregates after the withdrawal of acoustic fields. As a result, mostly cell aggregates (>90%) mature into spheroids maintaining good cell viability. We further applied these acoustically assembled spheroids to drug testing to explore their potency in drug response. In conclusion, this 3D acoustic cell assembly device may pave the way for the scale-up fabrication of cell spheroids or even organoids, to enable flexible application in various biomedical applications, such as high-throughput screening, disease modeling, tissue engineering, and regenerative medicine.

Acoustofluidic Actuation of Living Cells

“…Extending this concept, Miao et al developed a vertical, multi-layered approach for BAW acoustofluidic cell cluster patterning, effectively creating a three-dimensional cell lattice and, thus, expanding the method's theoretical capacity (Figure 3C). If integrated with Luo's platform, this innovation could herald a new era of high-throughput, low-cost, automated spheroid generation equipment [106]. In parallel with the trajectory of 3D bioprinting, the prospect of commercially available acoustofluidic devices promises to significantly advance the field of biomedical engineering.…”

“…(C) Acoustic pressure field generated by a 3D acoustic assembly device, and suspended cells are periodically distributed in a 3D dot-matrix pattern. Reprinted with permission from[106]. (D) Acoustic cell patterning can be used to produce hyaline cartilage tissue models, maintaining deep zone cytoarchitecture.…”

Trends in Photopolymerizable Bioinks for 3D Bioprinting of Tumor Models

“…High throughput formation of spheroid can improve the rapid testing of drugs. Recently high throughput spheroid formation via acoustic assembly based device to form cellular aggregates in GelMA based bioink was explored for hepatocellular carcinoma spheroid formation and their retrieval for rapid drug screening (Figure a) . In another study, dot extrusion of pancreatic adeno carcinoma cells (PDAC) mixed with normal human dermal fibroblasts (NHDFs) was explored for heterospheroid formation in GelMA-based bioink for cell–ECM, cell–cell interactions, and drug screening (Figure b).…”

“…Recently high throughput spheroid formation via acoustic assembly based device to form cellular aggregates in GelMA based bioink was explored for hepatocellular carcinoma spheroid formation and their retrieval for rapid drug screening ( Figure 4 a). 112 In another study, dot extrusion of pancreatic adeno carcinoma cells (PDAC) mixed with normal human dermal fibroblasts (NHDFs) was explored for heterospheroid formation in GelMA-based bioink for cell–ECM, cell–cell interactions, and drug screening ( Figure 4 b). Recently, Yang et al created a multicellular tumor spheroid model by combining microfluidics and a DMD device with controllable cell ratios and spheroid sizes for anticancer drug screening.…”

“…(a) GelMA based C3A cell spheroid formation using a 3D acoustic assembly device followed by photopolymerization for cell aggregate stabilization and retrieval of spheroids after 3 days for drug testing. Reproduced with permission from ref ( 112 ) with license under CC BY 4.0 ( ). No changes were made to the copyrighted material.…”