Successful islet isolation from preserved rat pancreas following pancreatic ductal collagenase at the time of harvesting.
In an attempt to develop an improved method for preserving the pancreas prior to islet isolation, the effects of warm and cold ischemia were examined on rat pancreases, from which reproducible high yields of islets can be obtained when fresh. Both warm and cold preservation rapidly decreased islet yield. Use of Hanks' or modified Sacks' solution also led to marked decrease in islet yield. After 6 hrs of preservation, the islet yield was 1/5-1/10 of those of fresh pancreases (374 +/- 74, n = 14), and no islets were obtained after 24 hrs of preservation regardless of the preservation solution. Monitoring of ductal pressure during forced injection of Hanks' solution in 6 hrs-preserved pancreas with HBSS showed a significantly earlier and lower peak of pressure than those of fresh pancreases. On the other hand, simple hypothermic preservation after pancreatic ductal distention with collagenase Hanks' solution at the time of harvesting resulted in a significantly higher islet yield up to 6 hours (171 +/- 58, n = 14, P less than 0.01), as compared with conventional methods. The viability of the islets isolated by this method was confirmed by the ability to restore normoglycemia of STZ-induced diabetic B6 mice on transplantation of 400 islets in the renal subcapsular space. These findings indicated that loss of the integrity of the ductal system against forced collagenase injection during cold preservation led to poor distention and digestion of the pancreas, ductal collagenase injection at the time of pancreas harvesting followed by simple preservation is recommended to obtain viable islets from the preserved pancreas.
Gotoh
M
M
Second Department of Surgery, Osaka University Medical School, Japan.
Ohzato
H
H
Dono
K
K
Kawai
M
M
Yamamoto
H
H
Kanai
T
T
Monden
M
M
Mori
T
T
D000818:Animals; D002452:Cell Count; D002469:Cell Separation; D007267:Injections; D007511:Ischemia / Q000473:pathology; D016381:Islets of Langerhans Transplantation / Q000379:methods*; D003012:Microbial Collagenase; D009926:Organ Preservation; D010179:Pancreas / Q000098:blood supply / Q000166:cytology / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D013696:Temperature
D003012:Microbial Collagenase