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Abstract 3D cell cultures are rapidly emerging as a promising tool to model various human physiologies and pathologies by closely recapitulating key characteristics and functions of in vivo microenvironment. While high‐throughput 3D culture is readily available using multi‐well plates, assessing the internal microstructure of 3D cell cultures still remains extremely slow because of the manual, laborious, and time‐consuming histological procedures. Here, a 4D‐printed transformable tube array (TTA) using a shape‐memory polymer that enables massively parallel histological analysis of 3D cultures is presented. The interconnected TTA can be programmed to be expanded by 3.6 times of its printed dimension to match the size of a multi‐well plate, with the ability to restore its original dimension for transferring all cultures to a histology cassette in order. Being compatible with microtome sectioning, the TTA allows for parallel histology processing for the entire samples cultured in a multi‐well plate. The test result with human neural progenitor cell spheroids suggests a remarkable reduction in histology processing time by an order of magnitude. High‐throughput analysis of 3D cultures enabled by this TTA has great potential to further accelerate innovations in various 3D culture applications such as high‐throughput/content screening, drug discovery, disease modeling, and personalized medicine.
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This content will become publicly available on April 1, 2026
A Bioinert Hydrogel Framework for Precision 3D Cell Cultures: Advancing Automated High‐Content and High‐Throughput Drug Screening
Enhanced drug testing efficiency has driven the prominence of high‐content and high‐throughput screening (HCHTS) in drug discovery and development. However, traditional HCHTS in well‐plates often lack complexity of in vivo conditions. 3D cell cultures, like cellular spheroids/organoids, offer a promising alternative by replicating in vivo conditions and improving the reliability of drug responses. Integrating spheroids/organoids into HCHTS requires strategies to ensure uniform formation, systemic function, and compatibility with analysis techniques. This study introduces an easy‐to‐fabricate, low‐cost, safe, and scalable approach to create a bioinert hydrogel‐based inverted colloidal crystal (BhiCC) framework for uniform and high‐yield spheroid cultivation. Highly uniform alginate microgels are fabricated and assembled into a colloidal crystal template with controllable contact area, creating engineered void spaces and interconnecting channels within agarose‐based BhiCC through the template degradation by alginate lyase and buffer. This results in a multi‐layered iCC domain, enabling the generation of in‐vitro 3D culture models with over 1000 spheroids per well in a 96‐well plate. The unique hexagonal‐close‐packed geometry of iCC structure enables HCHTS through conventional plate reader analysis and fluorescent microscopy assisted by house‐developed automated data processing algorithm. This advancement offers promising applications in tissue engineering, disease modeling, and drug development in biomedical research.
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- PAR ID:
- 10585250
- Publisher / Repository:
- Wiley
- Date Published:
- Journal Name:
- Small Science
- Volume:
- 5
- Issue:
- 4
- ISSN:
- 2688-4046
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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