Insulin Secretion ELISA
Insulin secretion from hiPSC-derived pancreatic cells cultured on
scaffolds was assessed by ultrasensitive insulin ELISA kit (ALPCO
80-INSHUU-E01.1) at stages 4, 5 and 6 of differentiation (days 1, 5 and
10 post seeding in 96-well plates and every other day in bioreactors).
Following the manufacturer’s instructions, cell-culture supernatants,
standard solutions, and controls were added to anti-insulin pre-coated
microplate wells and incubated with detection antibody at room
temperature with shaking for 1 hour. Subsequently, the microplate was
washed six times with provided wash buffer and incubated with
3,3′,5,5′-Tetramethylbenzidine substrate for 30 minutes with shaking.
After addition of the stop solution, the absorbance of each sample was
measured at 450 nm.