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. 2021 Jan 20;11(2):e3889. doi: 10.21769/BioProtoc.3889

Table 2. Preparation of 3D matrices with varying mechanical properties.

Shown is a pipetting scheme for the preparation of collagen-fibronectin matrices with embedded cells. Volumes are shown for the generation of 1 ml of soft and stiff matrix (200 mM).

Collagen matrix calculation scheme Soft Stiff
Collagen stock solution 200 µl 200 µl
0.5M ribose solution (in medium) - 133.3 µl
leave on ice for 30 min
HEPES (stock = 1 M) 20 µl 20 µl
Fibronectin (stock = 1mg/ml) 20 µl 20 µl
1 M NaOH 4 µl 4 µl
Sodium bicarbonate (7.5% solution) 40 µl 40 µl
Phenol-red free medium (serum-free) 216 µl 82.7µl

mix collagen working solution

1:1 with cell suspension

Cells in medium 500 µl 500 µl
Final volume of collagen-cell mix 1000 µl