Page 276 - IJB-9-4

International Journal of Bioprinting 3D acoustically assembled cell spheroids with high-throughput

Figure 5. Retrieval of acoustically fabricated cell spheroids. (a) The first two bright images on the left show hundreds of spheroids released from the GelMA
hydrogel scaffold, and the image on the right shows immunofluorescence staining for cell nuclei (DAPI, blue) and albumin (ALB, green) in the spheroid.
(b) Spheroid size distribution. (c) Bright images of retrieved spheroid grown during a 1-week culture. (d) Bar chart of the spheroid diameter. (e) Live/dead
staining and (f) viability assessment of the retrieved spheroids. All scale bars: 100 μm each.

Figure 6. Anticancer target drug (sorafenib) testing on acoustically fabricated C3A spheroids. (a) Live/dead staining of C3A spheroids and 2D cultures
after 48 h treatment with sorafenib. (b) The cell viability was measured. The statistical analysis showed significant differences between the spheroids and
2D cultures for different drug concentrations. *P < 0.05. Scale bar: 100 μm.

concentration of sorafenib increased, showing a dose- creating levitated acoustic nodes (LANs) in three directions,
dependent response to the drug. Taken together, these forming a 3D dot-array (25 × 25 × 22) of LANs. Based
results demonstrated that our acoustically fabricated on this characteristic, a large number of cell aggregates
spheroids can be used to evaluate drug efficacy and have arranged with the LANs were successfully assembled. To
great potential for high-throughput drug screening. further improve the aggregate maturation, we adopted
a 3D GelMA hydrogel scaffold strategy to maintain the
structure of cell aggregates during the transfer process or
4. Conclusion incubation period. As a result, most cell aggregates (>90%)
In this work, we presented the ability to improve the matured into spheroids that can be easily retrieved from
production yield of cell spheroids using our novel 3D the GelMA hydrogel in a non-destructive manner. The
acoustic assembly device. This device was capable of acoustically fabricated spheroids also manifested a higher

Volume 9 Issue 4 (2023) 268 https://doi.org/10.18063/ijb.733