The 3-D Life ToGro Hydrogel Kit contains reagents for the preparation of hydrogels with a soft stiffness (400 Pa; shear modulus) and a RGD Peptide concentration of 0.4 mmol/l. The gel composition allows three-dimensional spreading and migration of most cells. The gel is formed by the crosslinking of modified dextran carrying the cell adhesion motif RGD (RGD-Dextran) and CD-Link. When the two reagents are combined, thiol groups on CD-Link form stable thioether bonds with SH-reactive groups on RGD-Dextran resulting in gel formation within 20 min. The formation of hydrogel is performed at physiological pH for optimal cell compatibility. CD-Link is composed of polyethylene glycol fused to a matrix metalloprotease (MMP)-cleavable peptide. The MMP-cleavable peptide is designed for a broad range of MMP cleavage including MMP1, MMP3, MMP7 and MMP9*. It allows cells to spread and migrate within the gel if they express the indicated MMPs.
3-D Life ToGro hydrogels can be dissolved by the addition of dextranase (3-D Life Dextranase Cat. No. D10-1), which allows recovery of chemically fixed or live cells for post-culture analyses (e.g. RT-PCR) or further cultivation.
For preparation of gels follow the instructions of the 3-D Life ToGro Protocol “3-D cell culture with 3-D Life ToGro Hydrogel”.
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