Insulin secretory characteristics of monkey pancreatic islets: a simple method of islet isolation and the effect of various density gradients on separation.
We describe a simple stationary digestion method of islet isolation and separation by various density gradients from monkey pancreas (Macaca radiata radiata). Effective method, different types and concentrations of collagenase were standardized. Sigma type XI collagenase yielded >1000 islets/gram pancreas at the concentration of 4 mg/ml and 3 ml Hank's/gram pancreas. Slow digestion with less concentration of collagenase was suitable for monkey islet isolation. Discontinuous density gradients of bovine serum albumin (BSA) and dextran were compared with standard Ficoll for separation of islets. Islet yield (1038 +/- 81), insulin secretory response (stimulation index, S.I.11) and histological examination revealed dextran gradients were more appropriate for monkey islets when compared to BSA and Ficoll. Insulin secretory characteristics of monkey islets were studied by exposing them to low and high concentrations glucose (S.I.11.5), arginine (S.I.4.2), leucine (S.I.2.3) and tolbutamide (S.I.1.7). The results indicated that the magnitude of glucose induced insulin secretion of monkey islet is about half as that of rat and mouse islets. However, it is higher than that of porcine and bovine islets. In conclusion, the knowledge of insulin secretory ability of Indian bonnet monkey islets together with the techniques of isolation and separation are useful tool for diabetic research especially islet transplantation.
Balamurugan
A N
AN
Departments of Physiology and Pathology, Christian Medical College and Hospital, Vellore 632 002, TN, India. balamuruganan@msx.upmc.edu
Ramakrishna
Banumathi
B
Gunasekaran
S
S
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient; D017364:Collagenases / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D007202:Indicators and Reagents; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008255:Macaca radiata; D014044:Tolbutamide / Q000494:pharmacology
D000596:Amino Acids; D007202:Indicators and Reagents; D007328:Insulin; D014044:Tolbutamide; D017364:Collagenases; D005947:Glucose