Islet isolation for allotransplantation: variables associated with successful islet yield and graft function.
OBJECTIVE
Efficient islet isolation is an important prerequisite for successful clinical islet transplantation. Although progressively improved, islet yield and quality are, however, unpredictable and variable and require standardisation.


METHODS
Since 1989 we have processed 437 pancreases using the automated method. The donor characteristics, pancreas procurement, and digestion and purification procedures including a wide enzyme characterisation of these pancreases were analysed and correlated with islet yield and transplant outcome.


RESULTS
By univariate analysis, islet yield was significantly associated with donor age (r=0.16; p=0.0009), BMI (r=0.19; p=0.0004), good pancreas condition (p=0.0031) and weight (r=0.15; p=0.0056), total collagenase activity (r=0.22; p=0.0001), adjusted collagenase activity/mg (r=0.18; p=0.0002), collagenase activity/solution volume (r=0.18; p=0.0002) and neutral protease activity/solution volume (r=0.14; p=0.0029). A statistically significant contribution to the variability of islet yield in a multivariate analysis performed on donor variables was found for donor BMI (p=0.0008). In a multivariate analysis performed on pancreas variables a contribution was found for pancreas weight (p=0.0064), and for a multivariate analysis performed on digestion variables we found a contribution for digestion time (p=0.0048) and total collagenase activity (p=0.0001). Twenty-four patients with type 1 diabetes received single islet preparations from single donors. In these patients, multivariate analyses showed that the reduction in insulin requirement was significantly associated with morphological aspects of islets (p=0.0010) and that 1-month C-peptide values were associated with islet purity (p=0.0071).


CONCLUSIONS
These data provide baseline donor, digestion and purification selection criteria for islet isolation using the automated method and indicate that the morphological aspect may be a clinically relevant measure of islets on which the decision for transplant can be based.
Nano
R
R
Medicine and Surgical Departments, S. Raffaele Scientific Institute, Via Olgettina 60, Milan, Italy. bertuzzi.federico@hsr.it
Clissi
B
B
Melzi
R
R
Calori
G
G
Maffi
P
P
Antonioli
B
B
Marzorati
S
S
Aldrighetti
L
L
Freschi
M
M
Grochowiecki
T
T
Socci
C
C
Secchi
A
A
Di Carlo
V
V
Bonifacio
E
E
Bertuzzi
F
F
D000293:Adolescent; D000328:Adult; D000368:Aged; D001331:Automation; D002102:Cadaver; D002423:Cause of Death; D002469:Cell Separation / Q000379:methods; D003922:Diabetes Mellitus, Type 1 / Q000601:surgery*; D005260:Female; D006323:Heart Arrest; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology*; D016381:Islets of Langerhans Transplantation / Q000502:physiology*; D008297:Male; D008875:Middle Aged; D010180:Pancreatectomy / Q000379:methods; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000379:methods; D014184:Transplantation, Homologous