A matrigel-free method to generate matured human cerebral organoids using 3D-Printed microwell arrays
Open accessAbstract
The current methods of generating human cerebral organoids rely excessively on the use of Matrigel or other external extracellular matrices (ECM) for cell micro-environmental modulation. Matrigel embedding is problematic for long-term culture and clinical applications due to high inconsistency and other limitations. In this study, we developed a novel microwell culture platform based on 3D printing. This platform, without using Matrigel or external signaling molecules (i.e., SMAD and Wnt inhibitors), successfully generated matured human cerebral organoids with robust formation of high-level features (i.e., wrinkling/folding, lumens, neuronal layers). The formation and timing were comparable or superior to the current Matrigel methods, yet with improved consistency. The effect of microwell geometries (curvature and resolution) and coating materials (i.e., mPEG, Lipidure, BSA) was studied, showing that mPEG outperformed all other coating materials, while curved-bottom microwells outperformed flat-bottom ones. In addition, high-resolution printing outperformed low-resolution printing by creating faithful, isotropically-shaped microwells. The trend of these effects was consistent across all developmental characteristics, including EB formation efficiency and sphericity, organoid size, wrinkling index, lumen size and thickness, and neuronal layer thickness. Overall, the microwell device that was mPEG-coated, high-resolution printed, and bottom curved demonstrated the highest efficacy in promoting organoid development. This platform provided a promising strategy for generating uniform and mature human cerebral organoids as an alternative to Matrigel/ECM-embedding methods.
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A critical factor affecting differentiation in 3D printed PDMS microwells was the ridges of the culture vessel, unlike previous studies using smooth wells.
“They found that a critical factor affecting differentiation was the ridges of the culture vessel, unlike previous studies using smooth wells.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [174]
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- Authors
- Chen, C., Rengarajan, V., Kjar, A., & Huang, Y.
- Affiliations
- Utah State University (US)
- Corresponding author
- Huang, Yu
- Venue
- Bioactive Materials
- Publisher
- Elsevier BV
- Published
- 2021-04
- Volume
- 6
- Issue
- 4
- Pages
- 1130-1139
- ISSN
- 2097-1192 (ISSN-L)
- 2452-199X
- PMC
- PMC7577195
- OpenAlex
- W3093993828
- Cited by
- 77
- References
- 85
- Citations by year
- 2026: 8 · 2025: 12 · 2024: 20 · 2023: 12 · 2022: 18 · 2021: 7
- FWCI
- 2.91
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Bioactive Materials (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Utah Agricultural Experiment Station
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Keywords
- brain organoids, organoid maturation, printing resolution
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