Dynamic culture of bioprinted liver tumor spheroids in a
pillar/perfusion plate for predictive screening of anticancer drugs
Abstract
Recent advancements in three-dimensional (3D) cell culture technologies,
such as cell spheroids, organoids, and 3D bioprinted tissue constructs,
have significantly improved the physiological relevance of in
vitro models. These models better mimic tissue structure and function,
closely emulating in vivo characteristics and enhancing
phenotypic analysis, critical for basic research and drug screening in
personalized cancer therapy. Despite their potential, current 3D cell
culture platforms face technical challenges, which include user
unfriendliness in long-term dynamic cell culture, incompatibility with
rapid cell encapsulation in biomimetic hydrogels, and low throughput for
compound screening. To address these issues, we developed a 144-pillar
plate with sidewalls and slits (144PillarPlate) and a complementary
144-perfusion plate with perfusion wells and reservoirs
(144PerfusionPlate) for dynamic 3D cell culture and predictive compound
screening. To accelerate biomimetic tissue formation, small Hep3B liver
tumor spheroids suspended in alginate were printed and encapsulated on
the 144PillarPlate rapidly by using microsolenoid valve-driven 3D
bioprinting technology. The microarray bioprinting technology enabled
precise and rapid loading of small spheroids in alginate on the pillar
plate, facilitating reproducible and scalable formation of large tumor
spheroids with minimal manual intervention. The bioprinted Hep3B
spheroids on the 144PillarPlate were dynamically cultured in the
144PerfusionPlate and tested with anticancer drugs to measure drug
effectiveness and determine the concentration required to inhibit 50%
of the cell viability (IC 50 value). The perfusion plate
enabled the convenient dynamic culture of tumor spheroids and
facilitated the dynamic testing of anticancer drugs with increased
sensitivity. It is envisioned that the integration of microarray
bioprinting of tumor spheroids onto the pillar plate, along with dynamic
3D cell culture in the perfusion plate, could more accurately replicate
tumor microenvironments. This advancement has the potential to enhance
the predictive drug screening process in personalized cancer therapy
significantly.