VitroGel® Dilution Solution

VitroGel Dilution Solution is a ready-to-use solution to be used with the VitroGel High Concentration hydrogels to adjust the hydrogel concentration for different hydrogel strength.

The solution is room temperature stable with a neutral pH, which maintains a good physiological condition for hydrogel formation and can accelerate the hydrogel formation in combination with the cell medium. The Dilution Solution can be used to prepare a thin gel coating plate or injectable hydrogel with the high concentrations hydrogels.

The VitroGel Dilution Solution is offered in two versions:
TYPE 1: Contains sucrose to maintain the best osmolarity.
TYPE 2: Sucrose free formulation for cells sensitive to the sugar level in the medium

Specifications

• Ready-to-use to adjust the concentration of VitroGel High Concentration hydrogels

• Neutral pH

• Room temperature stable

• Transparent

• Compatible with VitroGel system for 2D & 3D cell culture and hydrogel injection

• Ships and store at room temperature

• Size: 50 mL

Frequently Asked Questions

• How to prepare the cell suspension to mix the hydrogel? Shall I add serum? If cells cultured in a complete cell culture medium, which is supplement with 10% FBS or other critical supplements, please prepare the cell suspension using the following methods before mixing it with hydrogel solution.

1. Prepare the cell suspension with 2X concentration (e.g. 100K), and mix with 100% FBS at 1:1 (v/v) ratio to get 1X cell suspension (50K) with 50% FBS.

2. Mix the diluted hydrogel solution with the cell suspension from above at 4:1 (v/v) ratio to get the final cells in the hydrogel at 10K with 10% FBS supplement.

If serum plan is an important role in your traditional cell culture, it is also important for 2D coating and 3D culture. Adding serum supplements in the hydrogel and adjusting the final serum concentration to the target level would support cell growth in the hydrogel system.

• How do I adjust the hydrogel formation time?– If VitroGel needs to be diluted more than 1:3 ratio, a longer waiting time (20-30 min) may be needed for soft gel formation. Using a higher volume of cell culture medium for mixing would help to accelerate the process of hydrogel formation.- If the hydrogel solidifies too fast after mixing with culture medium (showing as small solid gel chunk), adjust the mixing ratio by using less cell culture medium. For example, if mixing 4 mL diluted hydrogel solution with 1 mL cell culture medium lead to the solid gel chuck (particles), then mixing 4 mL diluted hydrogel solution with 0.5-0.8 mL cell culture medium would help to solve the issue.- On the other hand, if the hydrogel formation is too slow, which may happen when using low hydrogel concentration at 1:3 or 1:4 dilution or using a cell culture medium with very low ionic concentration, adjust the mixing ratio by using more cell culture medium. For example, if mixing 4 mL diluted hydrogel solution with 1 mL cell culture medium lead to a slow hydrogel formation, then mixing 4 mL diluted hydrogel solution with 1.5-4 mL cell culture medium would help to solve the issue.

• How do I adjust the stiffness of the final hydrogel? The stiffness of the final hydrogel can be adjusted by diluting the hydrogel solution before mixing with cell culture media. Our VitroGel Dilution Solution can help to adjust the hydrogel concentration. Please read the “First-time User Note” to learn how to prepare different VitroGel dilutions. If you need a higher hydrogel stiffness than the original product, please contact us at support@thewellbio.com.

• Can I harvest cells from the hydrogel after 3D culture? Yes, the cells can be harvested after 2D coating or 3D culture by using the VitroGel Cell Recovery Solution. VitroGel™ Cell Recovery Solution is a ready-to-use, enzyme-free solution to harvest 2D or 3D cultured cells from hydrogel fast and safely. The solution is compatible with the VitroGel hydrogel system and can recover cells from VitroGel in 15 minutes. VitroGel Cell Recovery Solution is room temperature stable, has a neutral pH and works at 37 °C operating temperature. The solution can maintain high cell viability during the recovery process. Cells can be sub-culture in both 2D and 3D culture after recovery.

DATA AND REFERENCES

Figure 1. Hydrogel formation by using DI water or VitroGel Dilution Solution.

The hydrogels were prepared at 1:1 dilution then 4:1 mixing with DMEM medium. VitroGel Dilution Solution helps accelerate the hydrogel formation faster than DI water.

Figure 2. The elastic modulus of stable hydrogel at different dilution ratio with DI water and VitroGel Dilution Solution.

Figure 3. Cells maintain high viability in VitroGel system at 1:3 dilution with VitroGel Dilution Solution.