- Add 3X of your medium supplement (e.g., R-Spondin, Noggin, etc) to the cell suspension before mixing with hydrogel at a 2:1 ratio. This way, you can ensure the hydrogel supplement is at least 1X.āÆ (you can prepare a 5X supplement in cell suspension if you like to promote the initial cell survival and growth)
- Increase the initial survival rate of organoids by seeding as a small colony instead of single cells and add ROCK inhibitor (such as Thiazovivin)
- If you want to mix VitroGelĀ® with BME, please add 10-20% BME to the cell suspension before mixing with VitroGelĀ®.
- For crypt, you can dilute VitroGelĀ® with DI water at a 1:1 ratio before mixing with cell suspension; we found that crypts have lower gel strength.