ISSN:
1573-0603
Keywords:
Dithizone staining
;
Islet isolation
;
Neonatal piglet
;
Pancreatic islets
;
Static-perifusion assay
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary Pancreatic islet transplantation shows promise as a future method for the treatment of insulin-dependent diabetes mellitus. However, as with reports of whole organ transplants in man, there are likely to be too few human donor organs. One potential approach to try and circumvent this problem is to seek alternative sources of pancreatic tissue. We have studied and developed a technique for isolating functional islets from one-day-old piglets with this in mind. Twelve pancreata, mean weight 1.27±0.36 g, were retrieved aseptically. The islets were isolated using a collagenase digestion procedure. An average of 116,000±45,000 islets/g tissue were isolated from each pancreas. After 24–48 h of culturing, a membrane-boundary was formed around the islets. Dithizone (DTZ) staining revealed greater than 50% of the presumed islets contained at least 5–6 red stained cells (i.e. insulin-containing cells). The insulin content of islet-like structures containing 50%, 10%, and 0% of DTZ stained cells was measured and revealed 32.1, 10.8 and 2.9 µIU, respectively, of immuno-reactive-insulin. In static-perfusion assays, a biphasic insulin secretion was observed in response to increased glucose stimuli. Preliminary studies showed that these islets were able to reverse the hyperglycemic condition of streptozotocin-induced diabetic rats. The present study indicates that one-day-old piglet islets can be utilized as the source of functioning islet cells. Isolation of these cells is reproducible and there appears to be no advantage technically and economically in using fetal pig pancreatic tissue rather than tissue from neonatal piglets for the purpose of obtaining pancreatic islets.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01404839
Permalink
