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Microstructured multi-well plate for three-dimensional packed cell seeding and hepatocyte cell culture
Vasiliy N Goral1, Sam H Au2, Ronald A Faris1
1Science and Technology, Corning Incorporated , Corning, New York 14831-0001, USA.
Biomicrofluidics
|November 8, 2014
Summary
This study introduces a microstructured plate for 3D cell culture using centrifugation. It promotes tissue-like cell structures and function without matrices, restoring cell polarity.
Area of Science:
- Biotechnology
- Cell Biology
- Tissue Engineering
Background:
- Standard cell culture often fails to replicate in vivo 3D cellular architecture.
- Achieving and maintaining 3D tissue-like morphology and function in vitro is challenging.
- Existing methods may require animal-derived or synthetic matrices.
Purpose of the Study:
- To develop a novel microstructured multi-well plate for 3D cell culture.
- To enable high-density cell seeding and culture using standard centrifugation.
- To promote 3D tissue-like cellular morphology and function without exogenous matrices.
Main Methods:
- Utilized a microstructured multi-well plate with microchannels/ditches.
- Employed standard laboratory centrifugation for high-density cell packing.
- Optimized centrifugation parameters (force, time, orientation) and seeding density.
- Cultured primary human hepatocytes for 7 days.
Main Results:
- Microfeatures and centrifugation successfully induced 3D cellular structures.
- Optimized cell packing enhanced cell-cell interactions.
- Primary human hepatocytes formed cord-like structures resembling 3D tissue.
- Restoration of cell membrane polarity was achieved without matrices or coagulants.
Conclusions:
- The microstructured plate and centrifugation method effectively support 3D cell culture.
- This approach promotes native tissue-like cellular morphology and function in vitro.
- It offers a matrix-free alternative for creating 3D cell models.
