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Long-Term Stable Poly(acrylamide) Brush Modified Transparent Microwells for Cell Attachment Studies in 3D
Inga Lilge1, Siyu Jiang1, Holger Schönherr1
1Physical Chemistry I, Department of Chemistry and Biology and Research Center of Micro and Nanochemistry and Engineering (Cμ), University of Siegen, Adolf-Reichwein-Str. 2, 57076, Siegen, Germany.
Macromolecular Bioscience
|January 4, 2017
Summary
Researchers created 3D cell microenvironments using stable poly(acrylamide) brushes in microwells. Fibroblast and tumor cells survived for over a week, adhering to well bottoms and sidewalls, showing less cytoskeleton alignment than in 2D cultures.
Area of Science:
- Biomaterials Science
- Cell Biology
- Microfabrication
Background:
- Creating controlled 3D cell microenvironments is crucial for understanding cell behavior.
- Poly(acrylamide) brushes offer stable, passivating surfaces for cell culture applications.
- Microwell formats allow for high-throughput screening and controlled cell positioning.
Purpose of the Study:
- To fabricate 3D cell microenvironments using stable poly(acrylamide) brushes in microwells.
- To investigate the behavior and viability of fibroblast and pancreatic tumor cells within these 3D structures.
- To analyze cell adhesion and cytoskeleton organization in microwells of varying shapes and sizes compared to 2D patterns.
Main Methods:
- Fabrication of microwells using micromolding in capillaries and surface-initiated polymerization of acrylamide.
- Functionalization of microwells with fibronectin (FN) to promote cell adhesion.
- Culturing NIH 3T3 fibroblast and Patu 8988T pancreatic tumor cells for over one week in 3D microwells.
Main Results:
- NIH 3T3 fibroblast and Patu 8988T cells adhered and remained viable in FN-coated microwells for >1 week.
- Cells attached to both the bottom and sidewalls of the microwells, unlike in 2D patterns.
- Cytoskeleton alignment was less pronounced in 3D microwells compared to 2D substrates, irrespective of microwell geometry.
Conclusions:
- Stable poly(acrylamide) brushes enable the creation of versatile 3D cell microenvironments.
- The 3D microwell format supports long-term cell viability and alters cell morphology compared to 2D cultures.
- This platform facilitates the study of cell behavior in controlled 3D settings, relevant for tissue engineering and cancer research.
