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3D printing of sacrificial thioester elastomers using digital light processing for templating 3D organoid structures
Benjamin J Carberry1,2, John E Hergert3, F Max Yavitt1,2
1Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO 80303, United States of America.
Biofabrication
|August 11, 2021
Summary
Digital light processing (DLP) biofabrication rapidly creates 3D sacrificial thioester elastomer molds. These molds pattern soft hydrogels, enabling controlled growth of intestinal stem cells (ISCs) into organoid-like structures.
Area of Science:
- Biofabrication
- Biomaterials Engineering
- Stem Cell Biology
Background:
- Biofabrication enables templating of cellularly-relevant structural features for tissue engineering.
- Organoid development benefits from biochemical and biomechanical stimuli to guide stem cell assembly and differentiation.
- Previous methods like laser-scanning created 3D overhangs in hydrogels for intestinal organoid architectures.
Purpose of the Study:
- To develop a high-throughput, complementary method to laser-scanning for creating 3D patterned structures for organoid culture.
- To utilize digital light processing (DLP) to print sacrificial thioester functionalized poly(ethylene glycol) (PEG) elastomers.
- To demonstrate the utility of these patterned substrates for culturing intestinal stem cells (ISCs).
Main Methods:
- Photopolymerization of PEG thiol and PEG norbornene with internal thioester groups to create degradable elastomers.
- Digital light processing (DLP) printing to fabricate 3D sacrificial shapes with high resolution and overhang features.
- Degradation of thioester elastomers using 2-mercaptoethanol and subsequent casting into Matrigel to create patterned void spaces.
- Culture of intestinal stem cells (ISCs) on the patterned Matrigel to assess cell behavior and tissue formation.
Main Results:
- DLP successfully printed thioester elastomer arrays with critical dimensions of 37 ± 4µm and resolutions of 22 ± 5µm.
- Sacrificial molds with overhang structures as small as 50µm were fabricated in minutes.
- Degradable elastomers patterned Matrigel, creating void spaces that high fidelity replicated the mold features.
- Intestinal stem cells (ISCs) cultured on patterned matrices formed confluent monolayers conforming to the underlying 3D geometry.
Conclusions:
- DLP-printed sacrificial thioester elastomers offer a rapid and robust method for fabricating 3D organoid-sized features in soft substrates.
- This technique facilitates the creation of patterned tissue culture environments for studying epithelial geometry effects on stem cell growth and differentiation.
- The method provides a valuable tool for advancing organoid research and tissue engineering applications.
