A modified automated method for isolation of viable pancreatic islets in laboratory animals.
To achieve successful islet transplantation, a high viability is required. For this reason an automated method including two chambers: one for islets isolation and one for recirculation and collection was developed. Recently, we produced a modified version of this work by building a similar system of glass where marbles were not used for disaggregation, and the pancreatic tissue had to be disrupted mechanically before the digestion phase. By using the reconfigured system, we obtained 260 +/- 20 islets from each Wistar albino rat (weighing 220 to 240 g) pancreas. Islets were observed at 35 minutes after the start of perfusion (closed circuit) and the optimum time to stop the isolation determined to be 40 minutes based upon islets viability. Although the present system is configured for islet isolation from small laboratory animals (rat, mouse), we have also obtained thousands of islets at 25 minutes after treatment of a 0.5-g piece of pig pancreas. Compared to the time-consuming manual method usually used for islet isolation from small laboratory animals, the new technique is economic, easy to use, and does not reduce islets viability.
Gürol
A O
AO
Istanbul University, Institute for Experimental Medical Research, Department of Immunology, Istanbul, Turkey. ogurol@yahoo.com
Yillar
G
G
Kurşun
A O
AO
Küçük
M
M
Deniz
G
G
Aktas
E
E
Oncan
N
N
Yilmaz
M T
MT
D000818:Animals; D001331:Automation; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D002470:Cell Survival; D007515:Islets of Langerhans / Q000166:cytology*; D023421:Models, Animal; D051381:Rats; D017208:Rats, Wistar