Isolation of Pancreatic Acinar Cells

Acinar cells were isolated as previously described [2 (link)]. Briefly, mice or rats (Charles River Laboratories, Wilmington, MA) were euthanized by CO₂ inhalation. The pancreas was minced in buffer-A [10 mM HEPES (pH 7.4), 95 mM NaCl, 4.7 mM KCl, 0.6 mM MgCl₂, 1mM NaH₂PO₄, 10mM glucose, 2mM glutamine, 0.1% bovine serum albumin, and 1× MEM amino acids (GIBCO-BRL, San Jose, CA)] and washed three times. Cells were then digested for 1h at 37°C in buffer-A containing 50 U/ml of type IV collagenase (Worthington, Freehold, NJ) with sustained shaking. The digest was filtered through a 200 μm mesh (Sefar American, Depew, NY), and the resulting groups of acinar cells were distributed in a 24-well Falcon tissue culture plate (Becton Dickinson, Franklin Lakes, NJ) and placed in a water bath shaking at 90-rpm under constant oxygen flow to recover. Human pancreatic acinar cells were prepared as previously described[17 (link)].