• How long before spheroid formation takes place in hydrogel?

  Normally, spheroid formation requires about 3-10 days after culture in the hydrogel system. The formation time may vary depending on different cell types. The small cell aggregation (colony formation) might happen overnight.Some readers

• How can I use VitroGel for co-culture or sandwich culture (layer by layer)?

  The co-culture of two or more different cell types can be performed with the VitroGel system. Besides adding different cell types together with the hydrogel solution for co-culture, each cell type can also be mixed with the hydrogel solution and be added layer by layer. After the first layer of hydrogel become stable, carefully overlay the second layer of cells/hydrogel mixture on top of the first layer of cells/hydrogel.Few readers

• How often does the cover media need to be changed for 3D culture?

  Typically, we recommend changing the cover medium every other day, similar to regular 2D cell culture. However, it depends on the experiment’s needs. Some experiments might require changing every 24 hours and some might not require changing for an entire week. Please contact us at support@thewellbio.com if you are unsure of this.Few readers

• How do I prepare the cell suspension to mix the hydrogel? Should I add serum?

  Cells can be prepared in regular complete cell culture medium and be directly mixed with VitroGel. If you want to adjust the concentration of the serum or other critical supplement in the final hydrogel, follow the steps below: If cells cultured in 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. Prepare the cell suspension with 2X concentratioFew readers

• Is it possible to cover cell spheroids with VitroGel and apply differentiation media on top?

  Yes, the medium on top can penetrate through the hydrogel.Few readers

• Shall I change the full amount of the cover medium or just partially?

  Changing 100% of the cover medium might cause the disruption of the hydrogel. We recommend adding additional fresh medium without removing the top medium for the first medium change. Afterward, change 50-80% of the cover medium. (Please check the recommended volume of additional cover medium in Table below.)Few readers

• What cell seeding density should I use?

  The final cell concentration can be optimized based on different cell types. We recommend preparing cell suspension at the following concentration for: 3D cell culture:5-2 x 106 cells/mL (the cell suspension needs to mix with VitroGel solution at the 4:1 ratio (VitroGel solution: Cell suspension at 4:1 v/v), which make the final cell concentration in the hydrogel 1-4 x 105 cells/mL) 2D hydrogel coating: 1-5 x 105 cells/mLFew readers

• Can cells move within the hydrogel matrix (migration/invasion)?

  Yes, the mobility of cells on VitroGel can be observed in both 3D culture and 2D hydrogel coating culture. Spheroid invasion assay, wound healing assay, or 3D cell migration can be performed with VitroGel.Few readers

• Can cells grown in the VitroGel be sub-cultured?

  Yes. Cells can be harvested from the VitroGel system by using the VitroGel Cell Recovery Solution and be sub-cultured for an additional period by using fresh VitroGel.Few readers