Instructions for making 0.6 wt% I2959 stock solution:
Add 50 ml of dH2O to 0.3 g I2959 in 50 ml conical vial
Heating to 60oC for a few hours may be necessary to dissolve I2959
Wrap vial completely in foil or opaque tape and store @ room temperature
Shelf life of a couple of years
Instructions for making macromer solutions:
Y = g dH2O ~ ml dH2O or = g PBS ~ ml PBS
I = g I2959 stock solution ~ ml I2959 stock solution
X = g macromer
W = weight fraction of macromer in polymerization solution
For 0.05 wt% final initiator concentration add the following amounts of Y & I to a measured mass of X:
Preparation of cells: Spin down cells for 10-12 minutes at 1000-1200 rpm and remove as much media or buffer from cells as possible. Filter macromer solution through a 20 micron filter into conical vial containing cells and resuspend cells by pipetting up and down.
Polymerize for 10 min at ~10 mW/cm2 (360 nm light, time can be adjusted for more or less intense light source)
Example: Using 0.1 g of PEG-DM to make a 10 wt% hydrogel
X = 0.1 g
W = 0.1
Y = 0.1 g (11-12*(0.1))/(12*0.1) = 0.817 ~ 0.817 ml
I = (1/11)*(X + Y) = 0.083 g ~ 0.083 ml
Total solution volume ~ 0.9 ml
Polymerize under 360 nm light source