Successful pancreas preservation by a perfluorocarbon-based one-layer method for subsequent pig islet isolation.
BACKGROUND
The oxygenation of human pancreas by the two-layer method (TLM) during cold storage was recently established for clinical islet transplantation. Simplification of TLM would facilitate the application of perfluorocarbon (PFC) as a regularly used preservation solution for subsequent islet transplantation. The present study examined whether PFC can be used in a one-layer method (OLM) for long-term pancreas preservation before isolation of adult pig islets.


METHODS
Resected pancreases were intraductally flushed with cold University of Wisconsin solution and immediately processed (n=6) or subjected to 7-hour storage by OLM (n=8) or TLM (n=10). Subsequently, pancreases were intraductally distended with collagenase NB-8 supplemented with neutral protease. Isolation and purification were performed as previously described.


RESULTS
Compared with unstored pancreases (3,670+/-740 islet equivalents [IEQ]) purified islet yield in TLM-stored organs (2,080+/-290 IEQ, P<0.05) was significantly decreased in contrast with OLM-preserved pancreases (3,110+/-520 IEQ, NS). No differences were observed between groups regarding purity (>90%), trypan-blue exclusion (>95%), adenosine triphosphate content, and mitochondrial viability of islets. Stimulation index during static glucose incubation (20 vs. 2.8 mm) was decreased after storage by TLM (1.81+/-0.20, P<0.05) but not by OLM (2.27+/-0.57) if compared with unstored pancreases (2.47+/-0.36). However, transplantation into diabetic nude mice resulted in sustained normoglycemia of recipients of either group until nephrectomy of graft-bearing kidneys was performed.


CONCLUSIONS
This study demonstrates that PFC alone can be used in a one-layer procedure for successful pig-pancreas preservation. This simplification can facilitate the broad application of PFC as pancreas preservation solution without reducing its benefits demonstrated by TLM.
Brandhorst
Daniel
D
Third Medical Department, University Hospital, Rodthohl 6, 35385 Giessen, Germany. Daniel.Brandhorst@innere.med.uni-giessen.de
Iken
Marcus
M
Brendel
Mathias D
MD
Bretzel
Reinhard G
RG
Brandhorst
Heide
H
D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005466:Fluorocarbons / Q000494:pharmacology*; D005978:Glutathione / Q000494:pharmacology; D007328:Insulin / Q000032:analysis / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology; D016381:Islets of Langerhans Transplantation; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology; D013552:Swine
D005466:Fluorocarbons; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose