Forskolin-stimulated adenylylcyclase activity: a marker to assess islet cell viability following cold storage in different solutions and to predict islet cell function following transplantation.
For clinical islet cell transplantation, short-term storage of islet cells is likely to be necessary, and it is imperative that the islet cells be kept as viable as possible during the period. However, there are little data on which preservative solutions are most suitable for the storage of islet cells after isolations or before transplantation. To estimate islet cell viability and transplantation success rate in the present study, adenylylcyclase activity was measured with a rapid new fluorometric assay in rat islet cells prior to transplantation, because cAMP plays an essential role in determining islet beta-cell viability and responsiveness to various hormonal stimuli. Adenylylcyclase activity was measured in islet cells stored for different periods of time 0, 3, 16, 24, 48, 96 h) and in different preservative solutions. Approximately 1,000 islet cells from each preservation group using University of Wisconsin (UW) solution were transplanted to streptozotocin-induced diabetes mellitus (DM) rats. Transplant success was evaluated by measuring blood glucose levels. Preoperative adenylylcyclase activity was compared with posttransplant islet cell function. The adenylylcyclase activity of UW solution was significantly higher than that of Euro-Collins solution and lactate-Ringer's solution through the different preservation time periods. Preoperative adenylylcyclase activity correlated well with posttransplant islet cell function in a rat model of DM. We conclude that adenylylcyclase activity can be used as a marker to assess islet cell viability as well as differences in preservation media and may predict islet cell transplant success.
Kanazawa
S
S
Department of Surgery, Mitsugi General Hospital, Hiroshima, Mitsugi-gun, Japan.
Tsunoda
T
T
Murakami
M
M
Iwamoto
S
S
Kimoto
M
M
Onuma
E
E
Majima
T
T
Yamamoto
Y
Y
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D015415:Biomarkers; D001786:Blood Glucose; D002470:Cell Survival / Q000502:physiology; D003080:Cold Temperature; D005576:Colforsin / Q000494:pharmacology*; D015925:Cryopreservation; D000242:Cyclic AMP / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism / Q000601:surgery; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006085:Graft Survival / Q000502:physiology*; D006982:Hypertonic Solutions / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology / Q000201:enzymology; D016381:Islets of Langerhans Transplantation; D007552:Isotonic Solutions / Q000494:pharmacology; D008297:Male; D019188:Organ Preservation Solutions / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D000077325:Ringer's Lactate; D013997:Time Factors
D015415:Biomarkers; D001786:Blood Glucose; C037096:Euro-Collins' solution; D006982:Hypertonic Solutions; D007552:Isotonic Solutions; D019188:Organ Preservation Solutions; D000077325:Ringer's Lactate; D005576:Colforsin; D000242:Cyclic AMP; D000262:Adenylyl Cyclases