Micro-ring structures stabilize microdroplets to enable long term spheroid culture in 384 hanging drop array plates
Open accessAbstract
Using stereolithography, 20 different structural variations comprised of millimeter diameter holes surrounded by trenches, plateaus, or micro-ring structures were prepared and tested for their ability to stably hold arrays of microliter sized droplets within the structures over an extended period of time. The micro-ring structures were the most effective in stabilizing droplets against mechanical and chemical perturbations. After confirming the importance of micro-ring structures using rapid prototyping, we developed an injection molding tool for mass production of polystyrene 3D cell culture plates with an array of 384 such micro-ring surrounded through-hole structures. These newly designed and injection molded polystyrene 384 hanging drop array plates with micro-rings were stable and robust against mechanical perturbations as well as surface fouling-facilitated droplet spreading making them capable of long term cell spheroid culture of up to 22 days within the droplet array. This is a significant improvement over previously reported 384 hanging drop array plates which are susceptible to small mechanical shocks and could not reliably maintain hanging drops for longer than a few days. With enhanced droplet stability, the hanging drop array plates with micro-ring structures provide better platforms and open up new opportunities for high-throughput preparation of microscale 3D cell constructs for drug screening and cell analysis.
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Hsiao et al. used these plates as a high throughput method for testing anticancer drug sensitivities on H9 human epithelial carcinoma cells.
“They continued on to use these plates as a high throughput method for testing anticancer drug sensitivities on H9 human epithelial carcinoma cells.”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [55]
The Materials and Methods section (Hanging Drop Formation and Spheroid Culture; page unavailable) confirms that Hsiao et al. used 384 hanging-drop plates to culture cancer-containing spheroids. It describes PC-3Luc GFP prostate cancer cells in a co-culture, not H9 cells, and the experiment assessed long-term culture rather than anticancer drug sensitivity. (LLM, conf=0.87)
Experimental — accuracy not yet measuredHsiao et al. designed a plate using micro-ring structures to stabilize drops, allowing 14-day culture of prostate cancer cells.
“To overcome this, one group, Hsiao et al. (55), designed a plate using micro-ring structures to stabilize the drops, allowing for 14day culture of prostate cancer cells.”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [55]
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- Authors
- Hsiao, A. Y., Tung, Y.-C., Kuo, C.-H., Mosadegh, B., Bedenis, R., Pienta, K. J., & Takayama, S.
- Affiliations
- University of Michigan (US)
- Corresponding author
- Takayama, Shuichi
- Venue
- Biomedical Microdevices
- Published
- 2011-11-05
- Volume
- 14
- Issue
- 2
- Pages
- 313-323
- ISSN
- 1387-2176 (ISSN-L)
- 1572-8781
- PMC
- PMC3304008
- OpenAlex
- W2110419119
- Cited by
- 129
- References
- 35
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 10 · 2023: 11 · 2022: 14 · 2021: 11 · 2020: 12 · 2019: 7 · 2018: 11 · 2017: 10 · 2016: 7 · 2015: 14 · 2014: 10 · 2013: 4 · 2012: 2
- FWCI
- 3.03
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Biomedical Microdevices (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Scholarworks@UNIST (Ulsan National Institute of Science and Technology) (submittedVersion)
- Funders
- National Cancer Institute (P50CA069568)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Engineering › Automotive Engineering › Additive Manufacturing and 3D Printing Technologies
- Keywords
- 3D cell culture, spheroid culture, stereolithography, injection molding, droplet spreading, surface fouling, drug screening
- MeSH terms
- Equipment Design, Humans, Spheroids, Cellular, Cell Culture Techniques, Cell Line, Tumor, High-Throughput Screening Assays
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