Pancreatic islet isolation variables in non-human primates (rhesus macaques).
BACKGROUND
Non-human primates (NHPs) are important preclinical models for pancreatic islet transplantation (PIT) because of their close phylogenetic and immunological relationship with humans. However, low availability of NHP tissue, long learning curves and prohibitive expenses constrain the consistency of isolated NHP islets for PIT studies. To advance preclinical studies, we attempted to identify key variables that consistently influence the quantity and quality of NHP islets.


METHODS
Seventy-two consecutive pancreatic islet isolations from rhesus macaques were reviewed retrospectively. A scaled down, semi-automated islet isolation method was used, and monkeys with streptozotocin-induced diabetes, weighing 3-7 kg, served as recipients for allotransplantation. We analysed the effects of 22 independent variables grouped as donor factors, surgical factors and isolation technique factors. Islet yields, success of isolation and transplantation results were used as quantitative and qualitative outcomes.


RESULTS
In the multivariate analysis, variables that significantly affected islet yield were the type of monkey, pancreas preservation, enzyme lot and volume of enzyme delivered. The variables associated with successful isolation were the enzyme lot and volume delivered. The transplant result was correlated with pancreas preservation, enzyme lot, endotoxin levels and COBE collection method.


CONCLUSIONS
Islet quantity and quality are highly variable between isolations. The data reviewed suggest that future NHP isolations should use bilayer preservation, infuse more than 80 ml of Liberase into the pancreas, collect non-fractioned tissue from the COBE, and strictly monitor for infection.
Andrades
P
P
Department of Surgery, University of Alabama at Birmingham, Birmingham, AL 35294-0019, USA. patoandrades@gmail.com
Asiedu
C K
CK
Gansuvd
B
B
Inusah
S
S
Goodwin
K J
KJ
Deckard
L A
LA
Jargal
U
U
Thomas
J M
JM
D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000473:pathology*; D004195:Disease Models, Animal; D005260:Female; D005978:Glutathione; D007328:Insulin; D007515:Islets of Langerhans / Q000166:cytology*; D016381:Islets of Langerhans Transplantation; D008253:Macaca mulatta; D008297:Male; D009924:Organ Culture Techniques / Q000379:methods* / Q000592:standards; D009926:Organ Preservation / Q000379:methods* / Q000592:standards; D019188:Organ Preservation Solutions; D011887:Raffinose
D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose