Comparative study of adult porcine pancreatic endocrine cell preparation using a technique of multiple injections and pancreatic duct cannulation without a proteolytic enzyme.
BACKGROUND
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.


OBJECTIVE
To compare two pancreatic injection methods: perfusion from an accessory pancreatic duct (cannulation method) and the traditional pancreatic tissue injection method (multiple injection method).


RESULTS
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).


CONCLUSIONS
These results indicate that pancreatic duct perfusion is useful for obtaining a high yield of PE cells from porcine pancreases.
Morimoto
Yoshihiko
Y
Division of Cell Replacement and Regenerative Medicine, Medical Research Institute, Tokyo Women's Medical University, Tokyo, Japan.
Nasu
Koko
K
Iwami
Yukiko
Y
Nagai
Kazutaka
K
Edamura
Kazuya
K
Ohgawara
Hisako
H
D000818:Animals; D002404:Catheterization; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D007267:Injections / Q000379:methods; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010183:Pancreatic Ducts; D010447:Peptide Hydrolases / Q000378:metabolism; D013552:Swine
D007328:Insulin; D010447:Peptide Hydrolases