Table 2.
Covalent Hydrogel Functionalization Approaches Tested and Their Outcomes After Seeding with Intestinal Epithelial Organoids
| Covalent hydrogel modification with peptides |
|
||
|---|---|---|---|
| Hydrogel | Peptide sequence | Outcome | |
| PEGDA | C-RKRLQVQLSIRT (C-AG73) | ** | |
| PEGDA | C-RGDS | ||
| PEGDA | C-RGDS + C-AG73 | * | |
| PEGDA | C-YIGSR | ** | |
| PEGDA | C-IKVAV | * | |
| PEGDA | C-RYVVLPR | − | |
| PEGDA | C-SIYITRF | * | |
| PEGDA | C-IAFQRN | * | |
| PEGDA | C-GEFYF | − | |
| Covalent hydrogel modification with ECM proteins | |||
| Hydrogel | Linking Chemistry | ECM Molecule* | Outcome |
| PEGDA | Sulfo-SANPAH | Collagen IV, Matrigel | ** |
| PEGDA | Acrylic acid-succinimidyl ester | Collagen IV, Matrigel | *** |
| PEGDA | 5-(acryloylamino)hexanoic acid-succinimidyl ester | Collagen IV, Matrigel | *** |
| PEG-norbornene | 2-iminothiolane | Collagen IV, Matrigel | *** |
| Controls | Outcome | ||
| Unmodified PEG hydrogel | − | ||
| Collagen IV-treated 96-well plate | ++++ | ||
| Outcome Key | |||
| − | Scattered dead cells, no cell aggregates | ||
| * | Sparse three-dimensional aggregates, no monolayers | ||
| ** | Scattered three-dimensional aggregates, no monolayers | ||
| *** | Dense three-dimensional aggregates, sparse areas with monolayer | ||
| ++++ | Confluent monolayer over entire area | ||
*
ECM molecules listed were each tested independently.