A Simple High-performance Modified Method For Isolating And Purifying The Rat Pancreatic Acinar Cells And Its Effects On The Functions Of The Cells

AIM To develop a simple high-performance and stabile modified method for isolating and purifying the rat pancreatic acinar cells ,then culture it, observe its morphology and to study its effects on the functions of the cellsMETHODS Eighteen SD rats were randomly divided into 2 groups, modified method group and intraductal perfusion method group(n=9). In modifled method group, the pancreatic tissues were obtained and shattered immediately after intraabdominal anesthesia, and then incubated in cell separating medium. The cell suspensions were obtained through two-time filtration and the pancreatic acinar cells were harvested after emundation in Hanks without Ca²⁺. In the other group, the pancreatic acinar cells were taken by intraductal perfusion method, The cell count, cell activities, purification rate, amylase in supernatant fluid and intracelular calcium concentration were assessed.RESULTS Freshly isolated acinar cells showed tendency to gather and form a monolayer without any extra cellular matrix and appeared as grape-like cell clumps and 4 hours later began to disperse uniformitily. Under light micrograph freshly isolated cells showed the typical ultrastructure of pancreatic acinar cells with zymogen granules (Z) and large amounts of rough endoplasmic reticulum (RER). When cells were cultured for 12 h, cultured acinar cells still exhibited Z granules and large amounts of RER . Some cellular debris was visible in the cultured cells, The cell count[(5.2±0.27)×10⁶/g ]and purification[(81.11±2.15)% of the pancreatic acinar cells harvested by the modified method had no statistical difference compared with those by the intraductal perfusion method[(4.8±0.49)×10⁶ / g,(80.89±3.98)% ,P>0.05].The cell activity [(91.33±1.80)% ] of modified method group was higher than that by the intraductal perfusion method (84±3)%, ,P<0.05. while the concentration of intracelular calcium [(3.71±0.42)μmol/L]was lower than that of intraductal perfusion method group[5.31±0.37,P<0.05].The amylase in supernatant fluid of modified method group is by turns (315.65±15.65)U/L, (836.84±52.69)U/L, (1621.25±158.58)U/L at 1h,4h and 10h , which was higher than that of the intraductal perfusion method , which is ( 283.92±19.45 ) U/L,( 621.71±42.34 ) U/L,(1406.06±151.38)U/L at the same time, P<0.05.CONCLUSION:Modified method is a simple,stable and effective method for isolating and purifying the pancreatic acinar cells with less affect on cell functions.