The 3-D Life Dextran-CD Hydrogel SG Kit provides reagents for setting up hydrogels for 3-D cell culture or related applications. Its major components are SG-Dextran and the crosslinker CD-Link. When the two reagents are combined, thiol groups on CD-Link form stable thioether bonds with SH-reactive groups on SG-Dextran. The resulting polymer network forms a hydrogel. The crosslinking step can be performed in the presence of cells to embed cells in the hydrogel.
Prior to the crosslinking step, cell adhesion peptides (e.g. 3-D Life RGD Peptide, Cat. No. 09-P-001) can be covalently attached to a portion of the SH-reactive groups on SG-Dextran to provide a cell-adhesive matrix.
The formation of hydrogels is performed at physiological pH for optimal cell compatibility. Gelation starts several minutes after mixing the components at room temperature (compare table in GP 2), providing enough time to conveniently manipulate the solution before the onset of gel formation.
A matrix metalloprotease (MMP)-cleavable peptide in CD-Link with a broad range for MMP cleavage including MMP1, MMP3, MMP7 and MMP9* allows cells to locally degrade the polymer network, if they produce the indicated MMPs. This allows cells to spread and migrate within the hydrogel. In most cases cell spreading and migration also requires the presence of adhesion peptides.
SG-Dextran hydrogels 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 after culture for post-culture analyses (e.g. RTPCR) or for further cultivation.