Description
The ToLuminate CD Photogel Kit provides reagents for setting up cell-compatible hydrogels formed by illumination with blue light (365-405 nm) on the basis of the thiol-ene chemistry. Its major components are norbornene-modified dextran (N-Dextran) and the thiol-modified crosslinker CD-Link. When the two reagents are combined and illuminated in the presence of LAP (Lithium phenyl-2,4,6-trimethylbenzoylphosphinate) thiol groups on CD-Link are activated and form stable thioether bonds with norbornene groups on N-Dextran. The components are mixed at physiological pH (pH 7.2) for optimal cell compatibility. There is ample time to place the pre-gel solution in any culture dish or container, since gelation starts only after illumination.
CD-Link contains a matrix metalloproteinase (MMP)-cleavable peptide sequence (Pro-Leu-Gly-Leu-Trp-Ala), which is cleaved by a broad range of MMPs including MMP1, MMP2, MMP3, MMP7, MMP9 and MMP13. It allows cells to locally degrade the polymer network, if they produce the indicated MMPs.
Cell adhesion peptides (e.g. 3-D Life RGD Peptide, Cat. No. 09-P-001) can be covalently attached to a portion of the norbornene groups on N-Dextran with the same chemical reaction as CD-Link to provide a cell-adhesive matrix.
3-D Life AgaFloat (Cat. No. A10-3) can be added to the pre-gel solution to avoid sedimentation of cells before illumination.
Dextran hydrogels crosslinked with CD-Link can be dissolved by the addition of dextranase (3-D Life Dextranase, Cat. No. D10-1), which allows the recovery of chemically fixed or live cells for post-culture analyses (e.g. RT-PCR) or for further cultivation.
For more information and instructions, please consult the General Protocol GP-5 "Preparation of ToLuminate Photogels", which may be downloaded below.
Application
- 3D cell cultures
- Bioprinting
Quantity
Up to 4 ml hydrogel.
Components
| Material | Quantity | Concentration of reactive groups | |
|---|---|---|---|
| N-Dextran |
600 µl | 30 mmol/L | MSDS |
| CD-Link, lyophilized | 3 x 200 µl* | 20 mmol/L* thiol groups | MSDS |
| 10x CB Buffer (pH 7.2) Phenol Red-free | 600 µl | n/a | |
| LAP, 10 mM | 600 µl | 10 mmol/L | MSDS |
| Water | 2 x 1500 µl | n/a | MSDS |
* Volume/concentration after reconstitution of lyophilizate.
Resources
- Product Datasheet (PDS): ToLuminate CD Photogel Kit
- Protocol: Preparation of ToLuminate Photogels
- Protocol: Dissolution of Dextran-Based Hydrogels with 3-D Life Dextranase (Recovery of Cells)
Related Products
Ilex Life Sciences LLC is an authorized distributor of Cellendes GmbH.