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Automated Nanodroplet Dispensing for Large-Scale Spheroid Generation via Hanging Drop and Parallelized Lossless
Viktoria Zieger1, Ellen Woehr2,3, Stefan Zimmermann1
1Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, D-79110 Freiburg, Germany.
Micromachines
|February 24, 2024
Summary
This study presents an automated workflow for generating and harvesting multicellular spheroids (MCSs) at scale. The high-throughput method ensures consistent spheroid quality and efficient recovery for advanced 3D cell culture applications.
Area of Science:
- Biotechnology
- Cell Biology
- Tissue Engineering
Background:
- Three-dimensional (3D) in vitro models, particularly multicellular spheroids (MCSs), are vital for studying complex biological processes like drug response and disease.
- Current methods for large-scale MCS production and recovery face challenges in consistency and efficiency.
Purpose of the Study:
- To develop and validate a high-throughput workflow for automated, large-scale production and parallel harvesting of multicellular spheroids.
- To address limitations in generating consistent MCSs and enable efficient transfer for downstream applications.
Main Methods:
- Utilized a hanging-drop technique with a non-contact dispenser for nanoliter droplet cell suspension dispensing.
- Implemented a centrifugation-based drop transfer method for spheroid harvesting into microtiter plates.
- Validated the workflow using MCF7 and HT29 cell lines for spheroid generation and post-transfer proliferation.
Main Results:
- Achieved 99.3% spheroid generation efficiency with a coefficient of variance below 8% for spheroid size consistency.
- Demonstrated 100% sample recovery during spheroid harvesting.
- Successfully cultured transferred HT29 spheroids in collagen matrices, showing continued proliferation.
Conclusions:
- The developed high-throughput workflow automates MCS production and harvesting, overcoming key limitations in current 3D cell culture techniques.
- This method facilitates prolonged spheroid cultivation, advanced assays, and reduces hands-off time, paving the way for more complex 3D cell culture protocols.
