Preparation of adult pig pancreatic cells: comparative study of methods with or without proteolytic enzymes.
Methods for the preparation of adult pig pancreatic cells using enzymatic- or auto-digestion are described, and a comparison of these methods was performed. A greater number of cells staining red with dithizone with the highest viability and insulin secretory response to glucose stimulation were obtained with dispersed pancreatic cells prepared by auto-digestion of the pancreas. On the other hand, many damaged cells were observed when the pancreas was exposed to collagenase or EDTA-Dispase and only a few cells stained red with dithizone. Insulin secretion in response to glucose stimulation was observed for up to 6 h in cells prepared using the auto-digestion procedure and an approximately 2- to 3-fold increase in insulin secretion was seen in response to glucose. Cells obtained by the collagenase or EDTA-Dispase exposure did not show a secretory response to either low (5.5 mmol/l) or high (16.7 mmol/l) glucose concentrations. This report shows that it is possible to prepare pancreatic endocrine cells from the adult pig pancreas which represents an inexpensive way to obtain cells for pancreatic cell transplantation or studies of their biochemical or physiological properties.
Ohgawara
H
H
Medical Research Institute, Tokyo Women's Medical College, Japan.
Kobayashi
A
A
Chong
S J
SJ
Akaike
T
T
Hashimoto
Y
Y
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D017364:Collagenases; D004492:Edetic Acid; D010450:Endopeptidases; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology; D010179:Pancreas / Q000166:cytology* / Q000502:physiology; D016035:Pancreas Transplantation; D013552:Swine; D014021:Tissue Preservation
D007328:Insulin; D004492:Edetic Acid; D010450:Endopeptidases; D017364:Collagenases; C023139:dispase