Redifferentiation of Expanded Human Islet Cells

Human islets were received 2–4 days following isolation from individual donors (Table 1). Islets were dissociated into single cells and cultured in CMRL 1066 medium containing 5.6 mM D-glucose and supplemented with 10% FCS (HyClone), 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml gentamycin, and 5 μg/ml amphotericine B (Biological Industries) (growth medium) as described [1 (link)]. The cultures were refed twice a week and split 1:2 once a week. For redifferentiation, expanded cells in passages 5–7 were trypsinized and seeded in ultra-low attachment plates with Redifferentiation Cocktail (RC) for 4–8 days as previously described [5 (link)]. The medium was replaced every two days.