Collagenase mtf c t

Islet isolation and transplantation were performed as previously described^{53 (link)}. Briefly, islets were isolated from monkey pancreas using the modified Ricordi method with collagenase MTF C/T (Roche, Mannheim, Germany). For the purification of isolated islets, the discontinuous Ficoll density gradient method was used. Next, purified islets were cultured with CMRL-1066 medium (Corning, NY, USA) containing 10% fetal bovine serum (FBS; Gibco-Thermo Fisher Scientific, Waltham, MA, USA) and 1% antibiotics (Gibco-Thermo Fisher Scientific) in a humidified 5% CO₂ atmosphere at 37 °C.

Resected partial or complete pancreases from the donor monkeys were processed as previously described to isolate the islets^{33 (link)}. Briefly, the islets were isolated from the pancreas using the modified Ricordi method with collagenase MTF C/T (ROCHE, Indianapolis, IN). The discontinuous Ficoll density gradient method was used to purify the islets. The purified islets were seeded with 30,000 IEQ per 150Ø Petri dish, and cultured with CMRL-1066 supplemented medium (CORNING, NY, USA) supplied with 10% fetal bovine serum (GIBCO-THERMO FISHER SCIENTIFIC, Waltham, MA, USA) and 1% antibiotics (GIBCO) in a humidified 5% CO₂ atmosphere at 37 °C. Cultured islets were used the next day (mean culture time was 20 (18–22) hours). On transplantation day, cultured islets were washed with fresh culture media, then counted for calculation of transplanted dose. Counted islets were suspended in CMRL 1066 transplantation medium (CORNING, NY, USA) without fetal bovine serum.