TrueGel3D™ is a chemically-defined hydrogel formed by mixing polymers with crosslinkers. This hydrogel enables the encapsulation of cells to permit their growth in a 3-dimensional environment to better mimic the native tissue environment.
The TrueGel3D™ hydrogel CD cell-degradable crosslinker and RGD peptide offer a predefined combination of reagents that allow cell growth in a 3D hydrogel with standard characteristics:
TrueGel3D™ hydrogel CD cell-degradable crosslinker and RGD peptide is recommended as a starting point for 3D hydrogel protocols when optimal application conditions have not yet been determined, or when optimization of gel firmness is not needed.
Follow instructions indicated in product datasheets for preparation of reagents.
To minimize oxidation of the thiol groups, do not expose CD cell-degradable crosslinker to air and room temperature longer than necessary. Make sure to close reagent caps immediately after each use.
Use culture medium, PBS or physiological solution to prepare your stock cell suspension or other biological sample.
IMPORTANT: please read the complete protocol before mixing your reagents.
Hydrogel mix preparation:
Your cell suspension should represent 1/5 of the final gel volume. Accordingly, the final concentration of cells in the complete gel will be 1/5 of your stock cell suspension.
Dissolving TrueGel3D™ hydrogel CD cell-degradable crosslinker and RGD peptide with TrueGel3D™ enzymatic cell recovery solution:
You can dissolve TrueGel3D™ Hydrogels containing live or chemically fixed cells by adding TrueGel3D™ enzymatic cell recovery solution to the culture medium. For example, a 30 μL gel can be dissolved with 300 μL of a 1:20 dilution of TrueGel3D™ enzymatic cell recovery solution in medium (30-60 minutes incubation, 37 °C). After dissolution of the gel, centrifuge the cell suspension and suspend the pelleted cells in fresh medium or physiological buffer. Repeat this washing procedure once or twice to remove remaining TrueGel3D™ enzymatic cell recovery solution. The removal of enzyme is important if cells are to be embedded again in dextran hydrogels to continue culture. If TrueGel3D™ enzymatic cell recovery solution is not removed completely, it can destabilize the newly set up hydrogel.