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 concentration (e.g. 100K) and mix with 100% FBS at 1:1 (v/v) ratio to get 1X cell suspension (e.g. 50K) with 50% FBS.
Mix VitroGel 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.
Note: Do not make 2X (or higher) concentration medium. The ionic molecules in the medium would affect hydrogel formation. The high concentration medium might make the gel chunky or give the gel a heterogeneous consistency.
The serum or critical supplement(s) in the hydrogel would affect cell growth, especially during the first 48 hrs. Please see the example figures below.
Bone marrow cells (OP9) 3D cultured in VitroGel LDP3 with 2% and 10% FBS. The cells were encapsulated in hydrogel matrix with 2% and 10% final FBS concentrations respectively. The images were taken 18 hours after cell seeding