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An Efficient Organoid Cutting Method for Long-Term Culture and High-Throughput Analyses.
Nicholas A Chartrain1,2, Marina V Pryzhkova3,4, Juliana I Candelaria3,4
14D Bio3 Center for Biotechnology and Department of Radiology and Bioengineering, Uniformed Services University of the Health Sciences, Bethesda, MD, 20814, USA.
Tissue Engineering and Regenerative Medicine
|June 16, 2025
Summary
This study introduces a 3D-printed jig for efficient organoid cutting, improving long-term culture viability and enabling high-throughput analysis for developmental and disease modeling.
Area of Science:
- Stem cell biology
- Biotechnology
- Regenerative medicine
Background:
- Human organoid models are crucial for research but face challenges in long-term maintenance due to nutrient and oxygen limitations.
- Current organoid sectioning methods are low-throughput and risk culture contamination, hindering advanced studies.
- Efficient organoid cutting is needed to overcome these limitations for sustained culture and high-throughput applications.
Purpose of the Study:
- To develop and optimize an efficient organoid cutting method using 3D printing.
- To enhance long-term organoid culture viability and enable high-throughput analyses.
- To facilitate consistent sample preparation for various organoid research applications.
Main Methods:
- Fabrication and optimization of four classes of 3D-printed organoid cutting jigs with blade guides.
- Culturing human pluripotent stem cell (hPSC)-derived organoids in mini-spin bioreactors and sectioning them every three weeks.
- Utilizing 3D-printed molds for embedding organoids in GelMA, Geltrex, or optimal cutting temperature compound (OCT) for array creation.
Main Results:
- All 3D-printed jigs facilitated rapid, uniform, and sterile organoid cutting, with a flat-bottom design showing superior efficiency.
- Organoid cutting improved nutrient diffusion, increased cell proliferation, and enhanced growth during long-term culture.
- 3D-printed molds enabled the creation of densely packed organoid arrays and consistent cryosections.
Conclusions:
- A novel 3D-printed organoid cutting and arraying method overcomes limitations in long-term culture and high-throughput processing.
- The simple, versatile design enhances organoid viability and sample preparation consistency.
- This approach supports improved organ development and disease modeling, drug screening, and single-cell spatial transcriptomics.
