How can I prevent some cells from being attached to the bottom resembling 2D coating when using VitroGel® for 3D cell culture?

Due to the homogeneous suspension of the cells with the VitroGel®, it is normal to see some cells attached to the bottom. However, to prevent this occurrence, a layer of hydrogel w/o cells at the bottom of the well plate can be first added before adding the gel/cell mixture. You can follow either method below to add a layer of hydrogel at the bottom:

Method 1:

1. Mix the VitroGel® 3D solution with VitroGel® Dilution Solution (Type 1 or Type 2) at 1:1 ratio

2. Add the mixture above to the bottom of the well plate and make sure the gel cover the whole bottom of the well

3. Keep the well plate at room temperature overnight or at 4 C for at least 2 hours. The coating plate is then ready for 3D cell culture application

Method 2:

1. Mix the VitroGel® 3D solution with VitroGel® Dilution Solution (Type 1 or Type 2) at 1:1 ratio

2. Mix the diluted hydrogel solution with basal cell culture medium at 4:1 ratio.

3. Add the mixture above to the bottom of the well plate and make sure the gel cover the whole bottom of the well

4. Keep the well plate at room temperature for 1 hour or at 4 C for 30 minutes. The coating plate is then ready for 3D cell culture application