Comparative Study of Adult Porcine Pancreatic Endocrine Cell Preparation Using a Technique of Multiple Injections and Pancreatic Duct Cannulation Without a Proteolytic Enzyme

Abstract

A method for isolation and primary monolayer culture of pancreatic endocrine (PE) cells from the porcine pancreas has already been established. It is very important for the PE cell preparation to expand the pancreas to separate PE cells from acinar cells. For this purpose, we developed a pancreatic injection system. To compare two pancreatic injection methods: perfusion from an accessory pancreatic duct (cannulation method) and the traditional pancreatic tissue injection method (multiple injection method). A comparison of the results of the two methods revealed that the PE cell yield was significantly higher with the cannulation method (2.97 +/- 0.59 x 10(7) cells per pancreas) than with the multiple injection method (0.89 +/- 0.15 x 10(7) cells per pancreas) (p < 0.0001). The number of dithizone-positive cells was significantly higher with the cannulation method (1.64 +/- 0.36 x 10(7) cells per pancreas) than with the multiple injection method (0.36 +/- 0.09 x 10(7) cells per pancreas) (p < 0.0001). The number of adhesion cells after 7 days of culture following isolation was higher with the cannulation method (1.07 +/- 0.26 x 10(7) cells per pancreas) than with the multiple injection method (0.36 +/- 0.03 x 10(7) cells per pancreas) (p < 0.0001). The glucose stimulation index of insulin secretion was higher with the cannulation method than with the multiple injection method (p < 0.01). These results indicate that pancreatic duct perfusion is useful for obtaining a high yield of PE cells from porcine pancreases.