Sigma type 5 colla genase

PPIs were isolated from pancreata of 8–15-day-old, pre-weaned Yorkshire piglets as previously described [44 (link)]. Briefly, pancreata were procured in less than 10 min and stored into cold HBSS (cold ischemic time < 1 h). Pancreata were minced into 1 mm³ pieces and digested with Sigma Type V Colla-genase (cat#C8051, Sigma Aldrich, Buchs, Switzerland) dissolved in HBSS (2.5 mg/mL) in 37 °C, 100 rpm shaking water bath for 15 min. After quenching with HBSS supplemented with 1% porcine serum (cat#26250084, Gibco, Waltham, MA, USA), digested tissues were filtered through a 500 μm metal mesh.

Animal procedures were authorized by the Institutional Animal Care and Use Committee (IACUC) at the University of California, Irvine. Pancreata from 8-to-15-day-old, pre-weaned Yorkshire pigs were procured for islet isolation [31 (link)]. All surgery and euthanasia were performed at University of California, Irvine. Euthanasia was done by cardiac puncture and exsanguination under anesthesia by IACUC approved personnel. Following procurement (<5 minutes), pancreata were placed in cold HBSS. The cold ischemic time was restricted to under 1 hour. Pancreata were then minced into 1mm³ tissue fragments and digested with Sigma Type V Collagenase (2.5 mg/mL, dissolved in HBSS; cat#C8051, Sigma Aldrich) in a 37°C, 100 rpm shaking water bath for 15 minutes. Enzymatic digestion was quenched with HBSS supplemented with 1% porcine serum solution, and the digested tissue was filtered through 500 μm stainless steel mesh.